CN104792770A - Method and device for water quality monitoring through continuous culture of luminous bacteria - Google Patents
Method and device for water quality monitoring through continuous culture of luminous bacteria Download PDFInfo
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- CN104792770A CN104792770A CN201510173288.7A CN201510173288A CN104792770A CN 104792770 A CN104792770 A CN 104792770A CN 201510173288 A CN201510173288 A CN 201510173288A CN 104792770 A CN104792770 A CN 104792770A
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Abstract
The invention discloses a method and device for water quality monitoring through continuous culture of luminous bacteria. The device comprises a luminous bacteria culture system, a reaction system and a detecting system, wherein the luminous bacteria culture system comprises a fermentation tank with a stirring device, a waste liquor tank and a material supplement tank; the liquid inlet of the fermentation tank is connected with the material supplement tank through a material supplement pump; the liquid outlet of the fermentation tank is connected with the waste liquor tank through a liquid drainage pump; a sampling opening is connected with the inlet of the reaction system through a sampling pump; the outlet of the reaction system is connected with the waste liquor tank; the detecting system wraps the periphery of the reaction system. The device is simple in structure, low in cost, convenient to operate and simple to maintain, can realize continuous automatic acute toxicity monitoring of water quality, and greatly simplifies monitoring steps. The sensitivity is equal to that of a luminous bacteria method of the national standard. Early warning is provided for sudden environmental accidents, so that corresponding measures are taken timely and high safety of water quality is ensured.
Description
Technical field
The present invention relates to the method and the device that utilize photogen Continuous Cultivation to carry out water quality monitoring.
Background technology
Photobacteria is a kind of bacterium that can be luminous.Its majority is Hai Sheng, is both with luminous planktonic organism the reason causing sea luminescence.Though photogen form is varied, physiological property is closely similar.Generally liquefaction is not produced to gelatin, after decomposing protein, do not form poisonous substance.
At present, domestic 3 kinds of conventional photobacterias are: photobacterium phosphoreum, Fermi operator, Qinghai Vibrion.Wherein, photobacterium phosphoreum is Gram-negative bacteria, can use in the mensuration of acute toxicity of water quality.Fermi operator is the Gram-negative bacteria of a kind of growth in ocean, is prevalent in marine environment and marine organism, is the pathogenic bacteria of some marine fishes.
Researchist finds when population density reaches certain threshold value, and Fermi operator can produce collective luminescence phenomenon; Research afterwards shows that this biological luminescence is because the accumulation of its self-induction agent causes; Fermi operator is exchanged mutually by this self-induction agent, starts the expression of related gene, thus causes the change of its phenotype.
Any inhibiting effect to cell metabolism mechanism, as various inorganic toxic material and organic poison, all can cause the minimizing of its luminescence, thus Fermi operator at present by general as environmental testing index.Qinghai Vibrion can launch blue green light (maximum emission wavelength 485nm) sustainedly and stably; once suffer from poisonous and harmful substance; very soon can suppressed luminescence; toxicity and the concentration of its luminous suppression degree and suffered poisonous and harmful substance have corresponding relation; and it is a kind of fresh water bacterium, so be widely used in potable water monitoring.In the world 20th century early eighties, bring into use photogen testing environment water pollution situation.China starts recommendation ocean photogen in nineteen ninety-five, as one of the biological method of ambient water quality pollution monitoring.
Above-mentioned photogen often occurs in environmental monitoring with the form of bacterium liquid, mycoderm or freeze-dried powder.The sharpest edges of photobacteria toxicity detection method be that it can Site Detection, method is fast, accurately, easy and simple to handle.
Start from 1940, Europe just has scholar to apply photobacteria to carry out toxicity detection to atmospheric pollution.In generation nineteen sixty, starts, and because the world is to the growing interest of environmental problem, uses the report of photobacteria toxicity detection to increase gradually.The 1970's Mos, just have the Biological Acute Toxicity detection method that scholar proposes based on photobacterium phosphoreum, Fermi operator, after this method is widely used.To the research of photobacteria this respect also comparatively early, red legend outstanding person waits people in a kind of photobacteria that can grow in freshwater lake of Late Cambrian in 1985, through years of researches by its called after Qinghai Vibrion, and is applied by this bacterium in China.The water quality toxicity that the nineties in 20th century, China proposed based on photobacterium phosphoreum detects national standard, and the application after this sending out bacterium expands gradually.
But when carrying out now Site Detection pollutant toxicity, the bacterium liquid after majority uses freeze-dried powder to recover, and photogen freeze-dried powder activity that is expensive, different batches there are differences on market, limits its application in on-line checkingi.
Summary of the invention
The object of this invention is to provide a kind of utilize photogen Continuous Cultivation to carry out water quality monitoring method and device, to overcome the deficiency having technology.
The described photogen Continuous Cultivation that utilizes carries out the device of water quality monitoring, comprises photogen culture systems, reactive system and detection system;
Described photogen culture systems comprises the fermentation tank, waste liquid tank and the feed supplement groove that are provided with stirring apparatus;
The inlet of described fermentation tank is connected with feed supplement groove by feed supplement pump, and leakage fluid dram is connected with waste liquid tank by positive displacement pump, and thief hatch is connected with the entrance of reactive system by sampling pump, and the outlet of reactive system (is connected with waste liquid tank; Described detection system is wrapped in described reactive system surrounding.
Adopt said apparatus, utilize photogen Continuous Cultivation to carry out the method for water quality monitoring, comprise the steps:
(1) frozen photobacterium phosphoreum is inoculated in the fermentation tank containing initial medium and cultivates;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system, add water quality sample to continuous print in reactive system simultaneously, detect luminous intensity by detection system, the water quality data of water quality sample can be obtained;
(3), after detection terminates, by rinse-system, the waste liquid in reaction tube is removed and rinse reaction tube, and detection next time can be carried out.
The invention has the beneficial effects as follows:
Acute toxicity of water quality on-Line Monitor Device structure of the present invention is simple, with low cost, easy to operate, safeguards simple; This method can realize water quality continuous and automatic Monitoring Acute Toxicity, and the Luminous bacteria of sensitivity and GB is suitable, simplifies monitoring step greatly.Using these apparatus and method to can be large-size lake river provides float type on-line continuous to monitor, and for outburst surroundings accident provides early warning to take corresponding measure timely, can greatly reduce the loss, and ensures water quality safety.
Accompanying drawing explanation
Fig. 1 is device schematic diagram.
Fig. 2 is that Continuous Cultivation bacterium liquid is to the response curve of zinc sulfate.
Fig. 3 is the active schematic diagram of the bacterium liquid cultivated.
Fig. 4 is the response curve of Continuous Cultivation bacterium liquid Pyrogentisinic Acid.
Embodiment
See Fig. 1, the described photogen Continuous Cultivation that utilizes carries out the device of water quality monitoring, comprises photogen culture systems, reactive system 2 and detection system 3;
Described photogen culture systems comprises:
Be provided with the fermentation tank 101 of stirring apparatus 102, waste liquid tank 103 and feed supplement groove 104;
Described fermentation tank 101 is closed container, inlet 107 is connected with feed supplement groove 104 by feed supplement pump 105, leakage fluid dram 108 is connected with waste liquid tank 103 by positive displacement pump 106, thief hatch 109 is connected by the entrance of sampling pump 110 with reactive system 2, and the outlet of reactive system is connected with waste liquid tank 103;
Preferably, described reactive system 2 is U-shaped reaction tube;
Described detection system 3 is wrapped in reactive system 2 surrounding, and preferably, described detection system 3 is the luminosity detector be made up of light activated element, be wrapped in described U-shaped reaction tube surrounding, during work, U-shaped reaction tube is in darkroom and detects its luminous intensity, and transfer digital signal display to;
Preferably, also comprise rinse-system, described rinse-system comprises flushing pump 401, abacus 402 and the sluice gun 403 that series winding connects, described sluice gun 403 is connected with the entrance of reactive system, after detection system working sample completes, rinsed reactive system by rinse-system, cleaning fluid enters waste liquid tank.
Adopt said apparatus, utilize photobacterium phosphoreum Continuous Cultivation to carry out the method for water quality monitoring, comprise the steps:
(1) be inoculated into by frozen photobacterium phosphoreum in the fermentation tank 101 containing initial medium, inoculum concentration is 1 ~ 2%, and temperature is 16 ~ 22 DEG C, rotating speed is cultivate 10 ~ 20 hours under 100 ~ 220r/min;
Bacterium liquid (in bacterium liquid, thalline the is in increased logarithmic phase) volume that inoculum concentration 2% refers to access nutrient culture media is 2% of culture volume;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
Fermentation tank 101 rotating speed is 100 ~ 220r/min, and temperature is 16 ~ 22 DEG C;
Feed rate gets dilutability D=0.08 ~ 0.12h
-1, preferred 0.1h
-1.
D=F/V (Feed flow rate is divided by tank volume), the flow velocity of representation unit volume nutrient solution, describes with dilution rate D, and what its represented is the renewal degree of material in tank.1/D represents the residence time of material in tank.Probably being equivalent to about one day time fills into the nutrient culture media equal with tank inner volume.
The component of initial medium and supplementing culture medium is:
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system, add water quality sample 5 to continuous print in reactive system simultaneously, detect luminous intensity by detection system, the water quality data of water quality sample can be obtained;
What step (2) obtained is 1: 1800 ~ 2200 containing the bacterium liquid of photobacterium phosphoreum and the volume ratio of water quality sample, preferably 1: 2000;
Described water quality sample is the water sample containing pollutant, and described pollutant is as zinc sulfate, phenol, potassium dichromate, comprehensive organism toxin;
(3), after detection terminates, by rinse-system, the waste liquid in reaction tube is removed and rinse reaction tube, and detection next time can be carried out.
Embodiment 1
Adopt the device of Fig. 1, utilize photogen Continuous Cultivation to carry out the method for water quality monitoring, comprise as follows:
(1) be inoculated into by frozen photobacterium phosphoreum in the fermentation tank 101 containing initial medium, inoculum concentration is 2%, and temperature is 18 DEG C, rotating speed is cultivate 16 hours under 200r/min;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
Fermentation tank 101 rotating speed is 200r/min, and temperature is 18 DEG C; Feed rate gets dilutability D=0.1h
-1;
Initial medium and supplementing culture medium are by following component and proportioning composition:
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system 2, add water quality sample to continuous print in reactive system 2 simultaneously, detect luminous intensity by detection system, the water quality data of water quality sample can be obtained;
What step (2) obtained is 1: 2000 containing the bacterium liquid of photobacterium phosphoreum and the volume ratio of water quality sample;
Described water quality sample is the water sample containing zinc sulfate;
Testing result is shown in Fig. 2.Fig. 2 is the TD-effect relation figure made according to zinc sulfate concentration and corresponding luminosity; As seen from Figure 2, the luminosity of working sample under different zinc sulfate concentration, and compare with blank group, obtain relative luminosity curve.As seen from the figure, R2=0.9887, show that curve linear is good, concentration and luminosity have linear relationship significantly.Survey relative rediance and can be used for characterizing water quality toxic concentration.
(3), after detection terminates, by rinse-system, the waste liquid in reaction tube is removed and rinse reaction tube, and detection next time can be carried out.
Embodiment 2
Adopt the device of Fig. 1, photogen Continuous Cultivation stability detected, comprises as follows:
(1) be inoculated into by frozen photobacterium phosphoreum in the fermentation tank 101 containing initial medium, inoculum concentration is 2%, and temperature is 18 DEG C, rotating speed is cultivate 18 hours under 200r/min;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
Initial medium and supplementing culture medium are by following component and proportioning composition:
Fermentation tank 101 rotating speed is 200r/min, and temperature is 18 DEG C;
Feed rate gets dilutability D=0.1h
-1;
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system 2, add dummy to continuous print in reactive system 2 simultaneously, detect luminous intensity by detection system, photogen activity data can be obtained;
What step (2) obtained is 1: 2000 containing the bacterium liquid of photobacterium phosphoreum and the volume ratio of dummy;
Described dummy is 3%NaCl solution.
Detection figure is shown in Fig. 3, wherein: OD represents optical density value, characterizes cell concentration; Luminosity represents luminosity, characterizes microbial activity; As seen from Figure 3: the bacterium liquid being undertaken cultivating by this device all can keep active in 160h, has the luminosity of enough carrying out toxicity detection, and in tank, photogen growth is stable, and activity difference is little.As can be seen here, this device can be used for long-term water quality comprehensive toxicity detection.
Embodiment 3
Adopt the device of Fig. 1, utilize photogen Continuous Cultivation to carry out the method for water quality monitoring, comprise as follows:
(1) be inoculated into by frozen photobacterium phosphoreum in the fermentation tank 101 containing initial medium, inoculum concentration is 2%, and temperature is 20 DEG C, rotating speed is cultivate 16 hours under 180r/min;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
Fermentation tank 101 rotating speed is 180r/min, and temperature is 20 DEG C;
Feed rate gets dilutability D=0.1h
-1;
Initial medium and supplementing culture medium are by following component and proportioning composition:
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system 2, add water quality sample to continuous print in reactive system 2 simultaneously, detect luminous intensity by detection system, the water quality data of water quality sample can be obtained;
What step (2) obtained is 1: 1800 containing the bacterium liquid of photobacterium phosphoreum and the volume ratio of water quality sample;
Described water quality sample is the water sample containing phenol;
Testing result is shown in Fig. 4.Fig. 4 is the TD-effect relation figure made according to phenol concentration and corresponding luminosity.As seen from Figure 4: the luminosity of working sample under different phenol concentration, and compare with blank group, obtain relative luminosity curve.As seen from the figure, R2=0.9698, show that curve linear is good, concentration and luminosity have linear relationship significantly.Survey relative rediance and can be used for characterizing water quality toxic concentration.
(3), after detection terminates, by rinse-system, the waste liquid in reaction tube is removed and rinse reaction tube, and detection next time can be carried out.
Claims (8)
1. utilize photogen Continuous Cultivation to carry out the device of water quality monitoring, it is characterized in that, comprise photogen culture systems, reactive system (2) and detection system (3);
Described photogen culture systems comprises the fermentation tank (101), waste liquid tank (103) and the feed supplement groove (104) that are provided with stirring apparatus (102);
The inlet (107) of described fermentation tank is connected with feed supplement groove (104) by feed supplement pump (105), leakage fluid dram (108) is connected with waste liquid tank (103) by positive displacement pump (106), thief hatch (109) is connected by the entrance of sampling pump (110) with reactive system (2), and the outlet of reactive system is connected with waste liquid tank (103);
Described detection system (3) is wrapped in described reactive system (2) surrounding.
2. device according to claim 1, it is characterized in that, also comprise rinse-system, described rinse-system comprises flushing pump (401), abacus (402) and the sluice gun (403) that series winding connects, and described sluice gun (403) is connected with the entrance of reactive system.
3. device according to claim 1 and 2, is characterized in that, described reactive system is U-shaped reaction tube.
4. device according to claim 3, is characterized in that, described detection system (3) is the luminosity detector be made up of light activated element, is wrapped in described U-shaped reaction tube surrounding.
5. adopt the device described in any one of Claims 1 to 4, utilize photobacterium phosphoreum Continuous Cultivation to carry out the method for water quality monitoring, it is characterized in that, comprise the steps:
(1) be inoculated into by frozen photobacterium phosphoreum in the fermentation tank 101 containing initial medium, inoculum concentration is 1 ~ 2%, and temperature is 16 ~ 22 DEG C, rotating speed is cultivate 10 ~ 20 hours under 100 ~ 220r/min;
(2) then add supplementing culture medium, carry out Continuous Cultivation, continuous print discharges the bacterium liquid containing photobacterium phosphoreum simultaneously, the volume of supplementing culture medium and the bacterium liquid phase etc. containing photobacterium phosphoreum of discharge;
(3) the bacterium liquid continuous print containing photobacterium phosphoreum step (2) obtained sends into reactive system, add water quality sample to continuous print in reactive system simultaneously, detect luminous intensity by detection system, the water quality data of water quality sample can be obtained.
(3), after detection terminates, by rinse-system, the waste liquid in reaction tube is removed and rinse reaction tube, and detection next time can be carried out.
6. method according to claim 5, is characterized in that, in step (2), fermentation tank 101 rotating speed is 100 ~ 220r/min, and temperature is 16 ~ 22 DEG C, and feed rate gets dilutability D=0.08 ~ 0.12h
-1.
7. method according to claim 5, is characterized in that, the component of initial medium and supplementing culture medium is:
8. method according to claim 5, is characterized in that, what step (2) obtained is 1: 200 ~ 2200 containing the bacterium liquid of photobacterium phosphoreum and the volume ratio of water quality sample.
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| CN113466216A (en) * | 2021-06-23 | 2021-10-01 | 同济大学 | Water quality monitoring method based on vibrio qinghai |
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Cited By (2)
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| CN108982889A (en) * | 2018-06-25 | 2018-12-11 | 中国科学院重庆绿色智能技术研究院 | A kind of synthetic biological toxicity in water on-line monitoring automatic analytical instrument |
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Inventor after: Feng Yaoyu Inventor after: Su Jiayuan Inventor after: Li Pei Inventor after: Meng Qin Inventor after: Hu Yue Inventor after: Jiang Yuanyuan Inventor after: Yao Hongli Inventor after: Wu Xiao Inventor after: Feng Yaoyu Li Pei Meng Su Jiayuan Qin Hu Yue Jiang Yuanyuan Yao Hongli Wu Xiao Yuan Xujun Inventor before: Feng Yaoyu Inventor before: Su Jiayuan Inventor before: Li Pei Inventor before: Meng Qin Inventor before: Hu Yue Inventor before: Feng Yaoyu Li Pei Meng Su Jiayuan Qin Hu Yue Jiang Yuanyuan |
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Effective date of registration: 20151008 Address after: 200237 Meilong Road, Shanghai, No. 130, No. Applicant after: East China University of Science and Technology Applicant after: Keyuan Electronic Tech Co., Ltd., Shanghai Address before: 200237 Meilong Road, Shanghai, No. 130, No. Applicant before: East China University of Science and Technology |
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Application publication date: 20150722 |