CN106148193A - Method for sampling and culturing algae cells in ambient air - Google Patents
Method for sampling and culturing algae cells in ambient air Download PDFInfo
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- CN106148193A CN106148193A CN201510173954.7A CN201510173954A CN106148193A CN 106148193 A CN106148193 A CN 106148193A CN 201510173954 A CN201510173954 A CN 201510173954A CN 106148193 A CN106148193 A CN 106148193A
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- 238000005070 sampling Methods 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 26
- 241000195493 Cryptophyta Species 0.000 title abstract description 7
- 238000012258 culturing Methods 0.000 title abstract 3
- 239000012080 ambient air Substances 0.000 title abstract 2
- 239000001963 growth medium Substances 0.000 claims abstract description 38
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 15
- 239000002609 medium Substances 0.000 claims abstract description 13
- 244000005700 microbiome Species 0.000 claims abstract description 13
- 238000005286 illumination Methods 0.000 claims description 21
- 229920001817 Agar Polymers 0.000 claims description 18
- 239000008272 agar Substances 0.000 claims description 18
- 239000000843 powder Substances 0.000 claims description 18
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 14
- 238000011534 incubation Methods 0.000 claims description 13
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 12
- 238000009835 boiling Methods 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 239000012530 fluid Substances 0.000 claims description 8
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 7
- 239000011736 potassium bicarbonate Substances 0.000 claims description 6
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 6
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 6
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 6
- 235000015497 potassium bicarbonate Nutrition 0.000 claims description 4
- 235000011181 potassium carbonates Nutrition 0.000 claims description 4
- 235000017550 sodium carbonate Nutrition 0.000 claims description 4
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- 230000003116 impacting effect Effects 0.000 claims 1
- 238000007789 sealing Methods 0.000 abstract description 10
- 239000007787 solid Substances 0.000 abstract description 8
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 238000012544 monitoring process Methods 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 2
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- 238000004113 cell culture Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 abstract 1
- 239000012531 culture fluid Substances 0.000 description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 12
- 239000000243 solution Substances 0.000 description 10
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 9
- 239000000443 aerosol Substances 0.000 description 9
- 229920001169 thermoplastic Polymers 0.000 description 9
- 239000004416 thermosoftening plastic Substances 0.000 description 9
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 239000007864 aqueous solution Substances 0.000 description 8
- 238000002156 mixing Methods 0.000 description 8
- 241000192660 Aphanizomenon Species 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 5
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 4
- 241000192608 Phormidium Species 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 4
- UFMZWBIQTDUYBN-UHFFFAOYSA-N cobalt dinitrate Chemical compound [Co+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O UFMZWBIQTDUYBN-UHFFFAOYSA-N 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 241000192584 Synechocystis Species 0.000 description 3
- 241000719329 Trentepohlia Species 0.000 description 3
- 235000009392 Vitis Nutrition 0.000 description 3
- 241000219095 Vitis Species 0.000 description 3
- 238000003915 air pollution Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 241000180279 Chlorococcum Species 0.000 description 2
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
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- 229910052927 chalcanthite Inorganic materials 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000007667 floating Methods 0.000 description 2
- 239000011565 manganese chloride Substances 0.000 description 2
- JKQOBWVOAYFWKG-UHFFFAOYSA-N molybdenum trioxide Chemical compound O=[Mo](=O)=O JKQOBWVOAYFWKG-UHFFFAOYSA-N 0.000 description 2
- 230000000243 photosynthetic effect Effects 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 235000011121 sodium hydroxide Nutrition 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- 239000011686 zinc sulphate Substances 0.000 description 2
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910018890 NaMoO4 Inorganic materials 0.000 description 1
- 241000192497 Oscillatoria Species 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
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- 210000000653 nervous system Anatomy 0.000 description 1
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the field of environmental air microorganism monitoring, in particular to a method for sampling and culturing algae cells in environmental air. The method comprises the following steps: 1) preparing a liquid culture medium; 2) adding an inorganic carbon source to the liquid medium; 3) preparing a solid culture medium, and subpackaging in culture dishes; 4) sampling algae cells in ambient air; 5) sealing the culture dish; and 6) culturing in a light incubator to obtain an algae cell culture sample in the air. The method provided by the invention has the advantage that the method can ensure that the collected algae cells can be cultured and grown under the laboratory condition.
Description
Technical field
The present invention relates to surrounding air microorganism monitoring field, in particular it relates to frustule sampling and the method cultivated in a kind of surrounding air.
Background technology
The fast development of economy and sharply increasing of population, promote the deterioration of the ecological environment, especially the pollution of urban environment air.Definition according to International Organization for Standardization, air pollution typically refers to owing to mankind's activity or natural process cause Cucumber to enter in air, present enough concentration, reach time enough, and therefore compromise comfortable, health and wellbeing or the phenomenon of environment of the mankind.In recent years, along with the aggravation of air pollution, the environmental consciousness of people is greatly improved, and associated research also has a lot.But just at present apparently, the most domestic focus primarily upon on Conventional pollution for air pollution focus of attention, such as: aerosolized particulate matter, reduced form pollutant (SO2, CO etc.), oxidized form pollutant (photochemical pollution etc.), petroleum type pollutant (NO2, alkene etc.) etc..But the concern of the aspects such as the secondary pollution polluted for surrounding air microorganism or have microorganism to participate in is relatively fewer, and correlational study is the most few.
Air microbe source complexity, drifts about along with air-flow again, it may be said that the most ubiquitous.Relatively large in solar radiation, easy dormancy or death under the higher environment of temperature.But suitably survive when condition, again can flourish again.According to the differences such as urban geography position, industrial structure, Population Capacity and urban ecological environment, air microbial contamination degree and primary pollution source the most difference.It addition, air ambient TSP concentration is relevant with airborne microorganisms content.The particularly particle size floating dust less than 10 m, the most portable microorganism is the most elegant.Overall suspended pellet can reduce ultraviolet and shine intensity, thus reduces the bactericidal action of ultraviolet.Air microbial contamination can not only cause the mankind and animals and plants transmission of disease, and industrial and agricultural products can be made putrid and deteriorated, directly threatens and affect health and the economic development of the mankind.As a example by frustule in microorganism, the microbial aerosol particle diameter that frustule is formed in atmosphere, typically at about 0.5 m, is mostly as air-flow drift and is diffused in air from soil with water body.Poisonous frustule also can discharge toxicant, even produces associating chemical reaction with other materials and makes toxicity further enhance, and have even can jeopardize health by respiratory system, nervous system etc..Accordingly, it would be desirable to the alga cells in air microbe is acquired, analyzes and monitors.
In the prior art, impingement method is generally used to realize the sampling of air microbe, the principle of the method is to utilize air extractor, with constant gas amount per minute, air is allowed to pass through narrow and small nozzle, make air and be suspended in microorganism particle therein formation high velocity air, when leaving nozzle, air-flow directive solid gathers face, gas turns round along collection face and leaves, granule is then pressed inertia and is continued straight ahead, clash into and adhere to and on collection face, capture microorganism particle, after cultivating, form bacterium colony, then detect.The method is mainly for microorganisms such as the heterotrophic bacteria in air, photosynthetic bacteria, fungus, virus, phagies, and lacks of sampling and the cultural method of alga cells in air.Different from these microorganisms, frustule is to have photosynthetic microbial body, its grown cultures mode and general cell, fungus, virus are entirely different, therefore, the air microbe sampling solid medium used at present and cultural method are not particularly suited for collection and the cultivation of alga cells in air.
The present invention relates to frustule sampling and the method cultivated in a kind of surrounding air, by environmental science relevant monitoring knowledge, set up frustule Sampling techniques in suitable air, the growing environment that simulation frustule is suitable, thus reach frustule sampling in air and the purpose cultivated.
Summary of the invention
It is an object of the invention to provide alga cells sampling and the method cultivated in a kind of surrounding air, with solve collection frustule cannot the problem of normal growth, realize the collection to active frustule floating in air, and make the active frustule gathered realize growth in laboratory conditions.
In order to achieve the above object, present invention employs techniques below step:
1) preparation fluid medium；
2) fluid medium prepared in step 1) adds inorganic carbon source, then with alkaline aqueous solution or acidic aqueous solution, pH value is adjusted to 7.1-7.2；
3) by step 2) gained fluid medium heats on electric furnace, and under slight boiling condition, add coagulator, make coagulator dissolve and mix；
4) step 3) gained solid medium is carried out autoclaving, be then sub-packed in culture dish, cover culture dish lid, be cooled to room temperature；
5) opening steps 4) culture dish lid after sterilizing, culture dish is put in air microbe impact sampler and sample；
6) culture dish after step 5) being sampled covers culture dish lid, and seals culture dish mouth；
7) being inverted in illumination box by the culture dish of step 6) and cultivate, cultivation temperature is 25-30 DEG C, daytime: night, intensity of illumination was 6000Lux-9000Lux, and incubation time is 6-20 days than for 20:4-8:16.
Further, algae described in step 1) is preferably blue-green alge and chlorella.
Further, preferred BG-11 or the BB culture medium of fluid medium described in step 1), BG-11(does not contains inorganic carbon source) culture fluid final concentration consists of: NaNO3 1500 mg/L、K2HPO4 40 mg/L、MgSO4·7H2O
75 mg/L、CaCl2·2H2O 36 mg/L, citric acid 6
Mg/L, ferric ammonium citrate 6 mg/L, Na2EDTA
1 mg/L、H3BO3 2.86 mg/L、MnCl2·4H2O
1.81 mg/L、ZnSO4·7H2O 0.222 mg/L、NaMoO4·2H2O 0.39 mg/L、CuSO4·5H2O 0.079 mg/L、Co(NO3)2·6H2O 0.0494 mg/L；BB culture fluid final concentration consists of: NaNO3 25 mg/L、CaCl2·2H2O
2.5 mg/L、MgSO4·7H2O 7.7 mg/L、K2HPO4
7.5 mg/L、KH2PO4 17.5 mg/L、NaCl 2.5 mg/L、Na2EDTA
50 mg/L、KOH 31 mg/L、FeSO4·7H2O
4.98 mg/L, dense H2SO4 1mL、H3BO3 11.42 mg/L、ZnSO4·7H2O
8.82 mg/L、MnCl2·4H2O 1.44 mg/L、MoO3
0.71 mg/L、CuSO4·5H2O 1.57 mg/L、Co(NO3)2·6H2O 0.49 mg/L。
Further, step 2) in add inorganic carbon source one of the most following: sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate.
Further, the final concentration of 5-20mg/L of carbanion, more preferably concentration are 10-15mg/L.
Further, step 2) described alkaline aqueous solution is preferably sodium hydrate aqueous solution or potassium hydroxide aqueous solution, more preferably molar concentration is the sodium hydrate aqueous solution of 1-2mol/L or molar concentration is the potassium hydroxide aqueous solution of 1-2mol/L, described acidic aqueous solution is preferably aqueous sulfuric acid that aqueous sulfuric acid or aqueous hydrochloric acid solution, more preferably molar concentration are 1-2mol/L or molar concentration is the aqueous hydrochloric acid solution of 1-2mol/L.
Further, in step 3), the preferred agar powder of coagulator of addition, addition is preferably 10-20g/L.
Further, in step 3), under the conditions of 121 DEG C, carry out autoclaving 20-30min.
Further, in step 4), solid matrix dispensed loading amount in culture dish is the 1/3-1/2 of culture dish volume.
Further, in step 5), air-flow sample rate is the preferred 25-32L/min of 20-35L/min(), acquisition time is 2-12 hour (preferably 5-8 hour).
Further, in step 6), culture dish mouth is sealed by the preferably self-styled sealing film of translucent thermoplastic.
Further, in step 7), preferred 28-30 DEG C of cultivation temperature, daytime: night than preferred 16:8-12:12, the preferred 7500-8500Lux of intensity of illumination, preferred 10-16 days of incubation time.
Compared with prior art, present invention have the advantage that
1) in the incubation of frustule long period, it is to avoid the loss of moisture in solid medium, and the substrate caused therefrom is dry and cracked, enables frustule to breed at cultured on solid medium；
2) in the incubation of frustule long period, a relatively moistening and airtight environment is provided for frustule growth, it is to avoid the interference of other microorganisms in air；
3) in the most airtight growing environment, solid culture based substrate can provide the nutrient needed for frustule, including inorganic carbon source；
4) the method has stable, simple and effective, economical and practical and is attainable feature under common experimental conditions, it is possible to gather and cultivate blue-green alge and the chlorella cell of abundant species, and cultivated frustule is easy to observe and separate.
Detailed description of the invention
Embodiment 1 : BG-11 (without inorganic carbon source) culture fluid adds27mg/LThe culture medium of sodium carbonate
The preparation BG-11 culture fluid without inorganic carbon source, is added thereto to Na2CO3, make the final concentration of 12mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.13.Culture medium is placed on electric furnace and heats, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 31L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 15 days than for 16:8.The 5th day cultivated at frustule, begins with small green spot and grows, and at the 13rd day, substantially presents green agglomerate, at the 15th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have the frustule of synechocystis, Aphanizomenon and Phormidium.
Embodiment 2 : BG-11 (without inorganic carbon source) culture fluid adds36mg/LThe culture medium of potassium carbonate
The preparation BG-11 culture fluid without inorganic carbon source, is added thereto to K2CO3, make the final concentration of 15mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.12.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 30L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 16 days than for 16:8.The 4th day cultivated at frustule, begins with small green spot and grows, and at the 11st day, substantially presents green agglomerate, at the 16th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have Chlorococcum, ball algae and the frustule of Phormidium.
Embodiment 3 : BG-11 (without inorganic carbon source) culture fluid adds24mg/LThe culture medium of sodium bicarbonate
The preparation BG-11 culture fluid without inorganic carbon source, is added thereto to NaHCO3, make the final concentration of 10mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.12.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 30L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 16 days than for 16:8.The 6th day cultivated at frustule, begins with small green spot and grows, and at the 12nd day, substantially presents green agglomerate, at the 16th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have the frustule of chlorella and synechocystis.
Embodiment 4 : BG-11 (without inorganic carbon source) culture fluid adds27mg/LThe culture medium of potassium bicarbonate
The preparation BG-11 culture fluid without inorganic carbon source, is added thereto to KHCO3, make the final concentration of 10mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.13.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 29L/min, and the sampling time is 5.5 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 16 days than for 16:8.Experimental result: the 5th day cultivated at frustule, began with small green spot and grows, at the 12nd day, substantially present green agglomerate, at the 16th day, culture dish is taken out, it is placed in basis of microscopic observation, records and take pictures, be found to have Aphanizomenon, Fructus Vitis viniferae Trentepohlia and the frustule of Oscillatoria.
Embodiment 5 :BBCulture fluid adds28mg/LThe culture medium of sodium carbonate
Preparation BB culture fluid, is added thereto to Na2CO3, make the final concentration of 15mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.12.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 30L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 15 days than for 16:8.The 4th day cultivated at frustule, begins with small green spot and grows, and at the 13rd day, substantially presents green agglomerate, at the 15th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have the frustule of chlorella, Aphanizomenon and Phormidium.
Embodiment 6 : BB Culture fluid adds35mg/LThe culture medium of potassium carbonate
Preparation BB culture fluid, is added thereto to K2CO3, make the final concentration of 10mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.11.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 30L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 16 days than for 16:8.The 5th day cultivated at frustule, begins with small green spot and grows, and at the 13rd day, substantially presents green agglomerate, at the 16th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have the frustule of Aphanizomenon and Phormidium.
Embodiment 7 : BB Culture fluid adds23mg/LThe culture medium of sodium bicarbonate
Preparation BB culture fluid, is added thereto to NaHCO3, make the final concentration of 10mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.13.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 30L/min, and the sampling time is 6 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 17 days than for 16:8.The 6th day cultivated at frustule, begins with small green spot and grows, and at the 12nd day, substantially presents green agglomerate, at the 17th day, is taken out by culture dish, be placed in basis of microscopic observation, record and take pictures, be found to have the frustule of Fructus Vitis viniferae Trentepohlia and synechocystis.
Embodiment 8 : BB Culture fluid adds26mg/LThe culture medium of potassium bicarbonate
Preparation BB culture fluid, is added thereto to KHCO3, make the final concentration of 14mg/L of carbonate, then with HCl solution, pH value be adjusted to 7.11.Culture medium is heated on electric furnace, agar powder is added under slight boiling condition, every L culture medium adds 18g, after agar powder dissolves mixing, while hot culture medium is sub-packed in culture dish, dispensed loading amount is the 1/2 of culture dish volume, covers culture dish lid, carries out autoclaving 30min under the conditions of 121 DEG C.After culture dish cools down, opening culture dish lid, culture dish is put into air microbe aerosol sampler and samples, air-flow sample rate is 29L/min, and the sampling time is 5.5 hours.Culture dish after sampling covers culture dish lid, and is sealed by culture dish mouth with translucent thermoplastic self-styled sealing film, makes air cannot be introduced in culture dish.Being put in illumination box by culture dish and cultivate, cultivation temperature is 30 DEG C, its daytime: night, intensity of illumination was 8000Lux, and incubation time is 16 days than for 16:8.The 5th day cultivated at frustule, began with small green spot and grows, at the 12nd day, substantially present green agglomerate, at 16 the days, culture dish is taken out, it is placed in basis of microscopic observation, records and take pictures, be found to have Aphanizomenon, Fructus Vitis viniferae Trentepohlia and the frustule of Chlorococcum.
Claims (7)
1. frustule sampling and the method cultivated in a surrounding air, it is characterised in that said method comprising the steps of:
(1) preparation fluid medium；
(2) in step (1) fluid medium, add inorganic carbon source, and to regulate solution ph be 7.1-7.2；
(3) fluid medium of step (2) is heated, under slight boiling condition, add coagulator；
(4) culture medium of step (3) is carried out autoclaving, be then sub-packed in culture dish, cover culture dish lid；
(5) the culture dish lid of opening steps (4) culture dish, carries out surrounding air frustule sample；
(6) culture dish of step (5) is covered culture dish lid, and culture dish mouth is sealed；
(7) culture dish of step (6) is inverted in illumination box cultivates.
2. the method for claim 1, it is characterised in that in step (1), fluid medium is one of following: the not BG-11 culture medium of containing sodium carbonate, BB culture medium.
3. method as claimed in claim 1 or 2, it is characterised in that in step (2), add inorganic carbon source be following one or more: sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate.
4. method as claimed in claim 3, it is characterised in that the final concentration of 5-20mg/L of carbanion.
5. the method for claim 1, it is characterised in that in step (3), added coagulator is agar powder, and addition is 10-20g/L.
6. the method for claim 1, it is characterised in that in step (5), uses impacting type air microorganism sampling device to sample, and air-flow sample rate is 20-35L/min, and acquisition time is 2-12 hour.
7. the method for claim 1, it is characterised in that in step (7) a metallic, cultivation temperature is 25-30 DEG C, daytime: night, intensity of illumination was 6000Lux-9000Lux, and incubation time is 6-20 days than for 20:4-8:16.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113355269A (en) * | 2020-09-24 | 2021-09-07 | 北京市园林科学研究院 | Low-temperature culture method and application of culture medium after inoculation in microbial experiment |
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| JPH0910785A (en) * | 1995-06-30 | 1997-01-14 | Pub Works Res Inst Ministry Of Constr | Control of bacteria of filter biological membrane |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH0910785A (en) * | 1995-06-30 | 1997-01-14 | Pub Works Res Inst Ministry Of Constr | Control of bacteria of filter biological membrane |
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| NORTH B A,ET AL: "Airbone algae from North-Centeral Florida USA", 《FLORIDA SCIENTIST》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113355269A (en) * | 2020-09-24 | 2021-09-07 | 北京市园林科学研究院 | Low-temperature culture method and application of culture medium after inoculation in microbial experiment |
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