CN206109061U - Collection born of same parents algae sewage treatment plant of bacillus blend system - Google Patents
Collection born of same parents algae sewage treatment plant of bacillus blend system Download PDFInfo
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- CN206109061U CN206109061U CN201620942763.2U CN201620942763U CN206109061U CN 206109061 U CN206109061 U CN 206109061U CN 201620942763 U CN201620942763 U CN 201620942763U CN 206109061 U CN206109061 U CN 206109061U
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Abstract
The utility model discloses a collection born of same parents algae sewage treatment plant of bacillus blend system mainly relates to little biomineralization and handles city domestic sewage field. The sterilization apparatus, microbial bioreactors, the mineral recovery unit that communicate in proper order including top -down, press bacteria and algae than 1 in the microbial bioreactors: 1000~1: 50 album born of the same parents algae and bacillus pack into in order to form the syntaxial system, the utility model has the advantages of is it through establishhing the fungus in the device the algae syntaxial system utilizes the collection born of the same parents algae to produce oxygen, and supply and balanced bacillus's growth demand has realized the application of microorganism sewage treatment in the large -scale production, has higher economic value and social effect.
Description
Technical field
This utility model is related to microbial mineralization and processes city domestic sewage field, and specifically cytoalgae-bacillus cereuss are common
The sewage-treatment plant of mixed system.
Background technology
With the continuous improvement flourished with people's living standard of China's industry, municipal sewage yield is increasing.
Due to the sewage treatment capacity wretched insufficiency of China, the municipal sewage for causing nearly 50% cannot get appropriate process and be discharged into river
The natural environment such as stream, lake, ocean, cause water body environment pollution, threaten ecological balance, and impact people's is healthy.And lead to
The focus that microorganism treating sewage is the research of sewage disposal in recent years is crossed, microorganism water purification is with its efficiency high, cost bottom, instant effect
Progressively become most promising water purification direction etc. advantage, now with the discussion to this kind of method and research gradually deeply, people
Gradually sight is concentrated on photosynthetic bacteria, bacillus cereuss, nitrobacteria, yeast, using mentioned microorganism as water purification
Primary body.Wherein, bacillus cereuss are the dominant populations in soil, with abundant protease, lipase, amylase, fiber
Plain enzyme etc., decomposition carbon, nitrogen system, phosphorus system, sulfur system pollutant that it can be strong, decomposing complicated polysaccharide, protein and water solublity has
Machine thing, becomes the focus of current water purification research.
However, above-mentioned conclusion major part is still within the discussion stage of laboratory, still it is short of corresponding in actual applications
Reaction treatment equipment, the economic worth for making achievement in research is greatly affected, and reduces the directive significance to actual production.
Additionally, in scale experiment is carried out, how the condition such as growth needs oxygen of bacillus cereuss is quickly cultivated and be protected
The concentration of bacillus cereuss and activity are held, just becomes one of main technical point for being produced using bacillus cereuss.If can reduce
Input to system, balances the growth demand of bacillus cereuss, maintains concentration and activity enough in microbial reaction, just becomes micro-
Biotechnology applications and the main direction of studying for producing.
Utility model content
The purpose of this utility model is the sewage-treatment plant for providing cytoalgae-bacillus cereuss co-mixing system, and it passes through
Bacterium-algae syntaxial system is set up in a device, using cytoalgae the growth demand of oxygen, supply and balance bacillus cereuss is produced, realize
Application of the microorganism sewage water process in large-scale production, with higher economic worth and social meaning.
This utility model for achieving the above object, is achieved through the following technical solutions:
The sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system, including the bactericidal unit being sequentially communicated from top to bottom,
Microorganism reactor, mineral recovery apparatus, are provided with cytoalgae and bacillus cereuss in the microorganism reactor.
The microorganism reactor includes the housing of transparent configuration, and on the side wall of the housing illuminator is provided with, described
Cytoalgae is scattered in the liquid phase in housing, the vertical reaction column for being provided with tubular, the inner chamber of the reaction column in the housing
In filled with embedding bacillus cereuss sodium alga acid capsule, the bottom of the reaction chamber is provided with aerator.
Device is communicated with the side wall of the housing, the linker is transparent configuration.
The housing is the cylindrical housings of lucite structure, and the reaction column is lucite cylinder structure, described
Illuminator is LED.
The aerator includes the air inlet pipe for being filled with sterilizing gas, and the outer end of the air inlet pipe is provided with intake valve
Door, the air inlet pipe is provided with coupled logical aeration tube, at the top of the aeration tube solarization air cap is provided with.
The bacillus cereuss are Bacillus licheniformis.
The bactericidal unit includes sterilization tank, and the asbestos heating plate for high temperature sterilize, institute are provided with the sterilization tank
Tell and be provided with the top of sterilization tank inlet, flow control valve is installed between the sterilization tank and microorganism reactor.
The layering dividing plate of several levels is provided with the sterilization tank, the layering dividing plate is relative with the section of sterilization tank
Should, one end of the sterilization tank is provided with breach, and the breach of the adjacent layering dividing plate is shifted to install, and the layering dividing plate is provided with
Several vertical baffle plates, the baffle plate is crisscross arranged to form serpentine channel.
The layering dividing plate is 2, and the baffle plate is 2.
The mineral recovery apparatus include reclaiming bucket, and the top for reclaiming bucket is provided with what is be connected with microorganism reactor
Discharge channel, is provided with discharge gate on the discharge channel, the side wall top of the recovery bucket is provided with discard solution discharge port, described
The bottom surface for reclaiming bucket is that taper is domatic, and placed in the middle being provided with the recovery bucket rotates the axis being connected, the axis with it
(15) bottom is provided with truss, and the bottom side of the truss is provided with and reclaims the material scraping plate that bucket is engaged, the bottom surface of the recovery bucket
Placed in the middle is provided with discharging opening, and on the discharging opening discharge valve is provided with.
Prior art is contrasted, the beneficial effects of the utility model are:
1st, this utility model provides a kind of device dedicated for processing city domestic sewage using microorganism, by micro- life
The achievement in research of thing water purification is applied to actual production, with higher operation instruction meaning and economic worth.The bactericidal unit
For carrying out sterilization treatment to handled sewage, found by using this device, bacterium algae ratio is pressed in the microorganism reactor
1:1000~1:50 loading cytoalgaes and bacillus cereuss can set up bacterium-algae syntaxial system, the spore under the conditions of aforementioned proportion
Bacillus growth is vigorous, and activity preferably, using cytoalgae oxygen is produced, and the growth for bacillus cereuss provides essential condition so as to meet dirty
The needs of water process, and reduce the input to cogeneration system, effectively save resource.Learnt by repetition test, using this practicality
New device and method to pass through precipitation filter, go carbonating city domestic sewage process after, can be by lung
The nitrogen and phosphorus pollution cycling of elements of edema is that the Institute of Micro-biology such as such as guanite produce Ore, that is, realize the purification of water quality, also can be become
Waste be changed into values, produces mineral and can be widely applied for the industries such as agricultural fertilizer, with prominent economic benefit, ecological benefits and society
Can benefit.
Compare through many experiments and find, 1 is compared by bacterium algae in the microorganism reactor:100 load cytoalgae and spore
Preferably, the growth of flora and activity are turning the growth conditions of the bacterium that bacillus is set up-algae syntaxial system in peak value to mineral
Metaplasia product aspect has prominent advantage.
The Bacillus licheniformis are harmless to environment and human body, and the metabolism to human body has regulatory function, also has
Have and be easy to by buying acquisition, the good advantage of biological filming performance is suitably applied sewage disposal.
The microorganism reactor is designed, the shell for culture and the sewage microbial reaction of bacterium-algae syntaxial system
The transparent configuration of body is engaged with the illuminator on the wall of side, and the foundation for bacterium-algae syntaxial system provides culture light conditions, described
The setting of linker, facilitates operator using the absorbance of liquid phase medium in the method detection housing of visible absorbance, and then
The cell density of cytoalgae is obtained, the judgement of running node is instructed.The bacillus cereuss are fixed on Sargassum by the method for embedding
In sour natrium capsule, fixation of the thalline in reaction column is realized, it is to avoid thalline is suspended in water, and realize recycling, while liquid
Only have cytoalgae in phase medium, the accurate detection to Synechocystis cell density can be realized, and then ensure to technique progress and behaviour
Make the accurate judgement of node, improve the success rate and high efficiency of wastewater treatment.By aerator aeration upwards, sewage is made micro-
Realize in bioreactor in order, effectively circulation, promote sewage and fixed bacillus cereuss carry out abundant, uniform contact,
Reaction, improves reaction efficiency.The disturbance of water body can also effectively be kept simultaneously, it is to avoid thalline is united, into ore deposit it is uneven the problems such as.It is described
Aerator it is simple for structure effectively, by being filled with the Ordered that aseptic gas are realized to water body, circulating effect is good.
It is that sewage water body fast and effectively sterilizes using the how empty heating dielectric structure of asbestos heating plate, is follow-up entrance
After bioreactor, the dominant microflora for keeping bacillus cereuss provides condition.The layering dividing plate is collectively forming horizontal three with baffle plate
The snakelike sterilization chamber of the baffling unhurried current of layer and the row of longitudinal direction three, enables sewage fully heat sterilization.
The mineral recovery apparatus mainly include reclaiming bucket and the truss with material scraping plate, by hang-up of the material scraping plate to bottom of struggling against
Agitaion enables handled waste water and the mineral of generation all to draw off, and the response rate of mineral is produced in raising.
The top discharge waste liquid being provided in mineral recovery apparatus of the overfall.Solid-liquid mixing after the completion of reaction
After thing is unloaded, bottom being gathered in solid product, liquid is flowed out on top by overfall, when mineral recovery apparatus reach saturation more
Afterwards, solid product is drawn off from the discharging opening of lower end, realizes the initial gross separation of solid-liquid, be conducive to the serialization operation for producing.
Description of the drawings
Accompanying drawing 1 is the growth curve of Bacillus licheniformis during this utility model embodiment 3 is tested;
Accompanying drawing 2 is cytoalgae growth curve during this utility model embodiment 3 is tested;
Accompanying drawing 3 is the growth curve of Bacillus licheniformis during this utility model embodiment 3 is tested;
Accompanying drawing 4 is the cytoalgae growth curve during this utility model embodiment 3 is tested in syntaxial system;
Accompanying drawing 5 is Ca during this utility model embodiment 3 is tested2+Concentration change figure;
Accompanying drawing 6 is the infrared light collection of illustrative plates of experiment and matched group Minerals during this utility model embodiment 3 is tested;
Accompanying drawing 7 is the infrared light collection of illustrative plates of experiment and matched group Minerals during this utility model embodiment 3 is tested;
Accompanying drawing 8 is the XRD spectrum of experimental group and matched group Minerals during this utility model embodiment 3 is tested;In figure:1. side
The monokaryon hydromagnesite of 4. fullerenes magnesite of solution stone 2. aragonite, 3. hydromagnesite 5.;
Accompanying drawing 9 is the XRD spectrum of experimental group and matched group Minerals during this utility model embodiment 3 is tested;In figure:1. side
The monokaryon hydromagnesite of 4. fullerenes magnesite of solution stone 2. aragonite, 3. hydromagnesite 5.;
Accompanying drawing 10 is the structural representation of the sewage-treatment plant of cytoalgae described in the utility model-bacillus cereuss co-mixing system
Figure;
Accompanying drawing 11 is the structural representation of layering dividing plate described in the utility model and baffle plate;
Accompanying drawing 12 is the schematic diagram of aerator described in the utility model.
Label shown in accompanying drawing:
1st, housing;2nd, linker;3rd, reaction column;4th, the sodium alga acid capsule of bacillus cereuss is embedded;5th, aerator;6th, enter
Trachea;7th, aeration tube;8th, solarization air cap;9th, sterilization tank;10th, inlet;11st, it is layered dividing plate;12nd, baffle plate;13rd, bucket is reclaimed;14th, unload
Material passage;15th, axis;16th, truss;17th, material scraping plate;18th, discharging opening;19th, breach;20th, discard solution discharge port.
Specific embodiment
With reference to specific embodiment, this utility model is expanded on further.It should be understood that these embodiments are merely to illustrate this
Utility model rather than restriction scope of the present utility model.In addition, it is to be understood that in the content for having read this utility model instruction
Afterwards, those skilled in the art can make various changes or modifications to this utility model, and these equivalent form of values equally fall within this Shen
Please limited range.
Involved instrument, reagent, material etc. in following embodiments, unless otherwise noted, are in prior art existing
Conventional instrument, reagent, material etc., can be obtained by regular commercial sources.Involved experimental technique in following embodiments, inspection
Survey method etc., unless otherwise noted, is existing normal experiment method, detection method etc. in prior art.
Embodiment 1:The sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system, agent structure include from top to bottom according to
The bactericidal unit of secondary connection, microorganism reactor, mineral recovery apparatus, the bactericidal unit includes sterilization tank 9, the sterilization tank
Asbestos heating plates for high temperature sterilize are installed, the top for telling sterilization tank 9 is provided with inlet 10 in 9, the sterilization tank 9 and
Flow control valve is installed between microorganism reactor.The layering dividing plate 11 of 2 levels is provided with the sterilization tank 9, it is described
Layering dividing plate 11 is corresponding with the section of sterilization tank 9, and one end of the sterilization tank 9 is provided with breach 19, the adjacent layering dividing plate
11 breach 19 is shifted to install, and the layering dividing plate 11 is provided with 2 vertical baffle plates 12, and the baffle plate 12 is crisscross arranged to be formed
Serpentine channel.The microorganism reactor includes the resin-case 1 of transparent configuration, and on the side wall of the housing 1 light source is provided with
Lamp, is communicated with device 2 on the side wall of the housing 1, the linker 2 is provided with transparent observation window, the housing 1 built with
Be inoculated with the BG-11 culture medium of cytoalgae, the vertical reaction column 3 for being provided with tubular in the housing 1, the reaction column 3 it is interior
Filled with the sodium alga acid capsule 4 of embedding bacillus cereuss in chamber, the microorganism reactor is interior to compare 1 by bacterium algae:100 load collection born of the same parents
, to form syntaxial system, the initial concentration of the cytoalgae is 0.35 × 10 for algae and bacillus cereuss9Individual/ml, the reaction chamber
Bottom is provided with aerator 5, and the aerator 5 is microporous aeration disc, the aeration plate with for being filled with aseptic gas
Equipment connects, and the mineral recovery apparatus include reclaiming bucket 13, and the top for reclaiming bucket 13 is provided with and microorganism reactor phase
The discharge channel 14 of connection, described recovery be provided with discard solution discharge port 20 at the top of 13 side walls of bucket, installs on the discharge channel 14
There is discharge gate, the bottom surface of the recovery bucket 13 is provided with discharge valve, the recovery bucket 13 and is provided with auger stripper.
Embodiment 2:The sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system, agent structure include from top to bottom according to
The bactericidal unit of secondary connection, microorganism reactor, mineral recovery apparatus, the bactericidal unit includes sterilization tank 9, the sterilization tank
Asbestos heating plates for high temperature sterilize are installed, the top for telling sterilization tank 9 is provided with inlet 10 in 9, the sterilization tank 9 and
Flow control valve is installed between microorganism reactor.The layering dividing plate 11 of 3 levels is provided with the sterilization tank 9, it is described
Layering dividing plate 11 is corresponding with the section of sterilization tank 9, and one end of the sterilization tank 9 is provided with breach 19, the adjacent layering dividing plate
11 breach 19 is shifted to install, and the layering dividing plate 11 is provided with 4 vertical baffle plates 12, and the baffle plate 12 is crisscross arranged to be formed
Serpentine channel.The microorganism reactor includes the lucite housing 1 of transparent configuration, is provided with the side wall of the housing 1
LED light source lamp, the side wall of the housing 1 is provided with the linker 2 of transparent organic glass structure, and the housing 1 is built with inoculation
There is the BG-11 culture medium of cytoalgae, the vertical reaction column 3 for being provided with tubular in the housing 1, in the inner chamber of the reaction column 3
Compare 1 by bacterium algae in sodium alga acid capsule 4 filled with embedding bacillus cereuss, the microorganism reactor:500 load cytoalgaes and
To form syntaxial system, the initial concentration of the cytoalgae is 0.50 × 10 to bacillus cereuss9Individual/ml, the bottom of the reaction chamber
Aerator 5 is installed, the aerator 5 includes the air inlet pipe 6 for being filled with sterilizing gas, the outer end of the air inlet pipe 6
Air intake valve is provided with, the air inlet pipe 6 is provided with coupled logical aeration tube 7, and the top of the aeration tube 7 is provided with solarization air cap
8, the mineral recovery apparatus include reclaiming bucket 13, and the top for reclaiming bucket 13 is provided with unloading of being connected with microorganism reactor
Material passage 14, described recovery be provided with discard solution discharge port 20 at the top of 13 side walls of bucket, and on the discharge channel 14 discharge valve is provided with
Door, the bottom surface for reclaiming bucket 13 is that taper is domatic, and placed in the middle being provided with the recovery bucket 13 rotates the axis being connected with it
15, the bottom of the axis 15 is provided with truss 16, and the bottom side of the truss 16 is provided with and reclaims the material scraping plate 17 that bucket 13 is engaged,
It is described reclaim bucket 13 bottom surface it is placed in the middle be provided with discharging opening 18, discharge valve is installed on the discharging opening 18.
When in use, need to load in the microorganism reactor after bacillus cereuss and cytoalgae, keep microorganism anti-
Answer temperature in device to be 28 DEG C, the use of intensity of illumination is 5000lx, the alternate illumination for carrying out illumination in 12 hours and 12 hours dark connects
Continuous culture 7 days;Then inlet 10 is opened, the city domestic sewage that precipitation is filtered, carbon elimination acidification is obtained will be passed through
Injection bactericidal unit, enters microorganism reactor after heat sterilization process, and holding intensity of illumination is 5000lx, carries out 12 little
The alternate illumination successive reaction of Shi Guangzhao and 12 hour dark 28 days, obtains waste liquid and mineral precipitation;Open mineral recovery apparatus
The waste liquid and mineral precipitation are entered, and collects mineral precipitation, that is, completed.
Embodiment 3:The induction that the bacterium-algae syntaxial system of different proportion is carried out under the conditions of Mg/Ca=6 is tested into ore deposit
Experiment purpose:The inducing culture condition that this experiment passes through setting Mg/Ca=6, using the cytoalgae of different proportion
Carry out mixing with Bacillus licheniformis setting up syntaxial system, for simulating cytoalgae described in the utility model-bacillus cereuss blending
Ore deposit reaction is cleaned in microorganism reactor in the sewage-treatment plant of system, so as to realizing to bacterium-algae syntaxial system most
The acquisition of good filling ratio, the determination of optimal performance technological parameter, and ore deposit effect is modeled to discussing by inducing culture
The feasibility that cytoalgae-Bacillus licheniformis described in the utility model set up syntaxial system, and bacterium-algae syntaxial system are demonstrate,proved to city
The purification of magnesium calcium and recycling ability in sanitary sewage, so as to verify effect of the present utility model.
1. strain
Bacillus licheniformis (Bacillus licheniformis SRB2), by laboratory isolation identification, are stored in refrigerator;
DNC wireless (Synechocysissp.PCC6803) is by Qingdao Institute of Biomass Energy and Bioprocess Technology of the Chinese Academy of Sciences
Present.
2. experiment reagent
Citric acid (Tianjin North Star Founder chemical reagent work);Ferric ammonium citrate (Shanghai fuzz chemical reagents corporation);Sodium nitrate
(Shanghai is strong along chemical reagents corporation);Dipotassium hydrogen phosphate (fine chemistry institute is recovered in Tianjin);Bitter salt (Tianjin
Bo Di Chemical Co., Ltd.s of city);CALCIUM CHLORIDE DIHYDRATE (Tianjin Kermel Chemical Reagent Co., Ltd.);Sodium carbonate (Tianjin
Rui Jin spy's chemical company);Boric acid (Tianjin is extensively into Chemical Co., Ltd.);(the rich Dihua work in Tianjin has four chloride hydrate manganese
Limit company);Zinc vitriol (Tianjin Bo Di Chemical Co., Ltd.s);(Jin Dui cities molybdenum industry science and technology is limited for two molybdic acid hydrate sodium
Responsible company);Copper sulfate pentahydrate (opens chemical reagents corporation in Tianjin);Cabaltous nitrate hexahydrate (Tianjin Rui Jin specialization
Product company);Anhydrous calcium chloride (opens chemical reagents corporation in Tianjin);(Chinese medicines group chemical reagent is limited for Magnesium dichloride hexahydrate
Company);Sodium hydroxide (Tianjin Bo Di Chemical Co., Ltd.s);Concentrated hydrochloric acid (Tianjin Ke Miou chemical reagents corporations);Beef
Leach powder (Beijing Luqiao Technology Co., Ltd.);Tryptone (Beijing bispin microbiological culture media products factory);Sodium Chloride
(the prosperous prosperous Chemical Co., Ltd. in Tianjin);Agar (Beijing Solartio Science & Technology CO, Ltd);
Sodium bicarbonate (Chinese BASF Chemical Co., Ltd.);Sodium carbonate (Tianjin Rui Jin spy Chemical Company);Anhydrous second
Alcohol (Laiyang economic and technological development zone Fine Chemical Works).
3. experimental apparatus
FC204 electronic analytical balances (Shanghai precision balance company limited);(Changzhou is lucky for XY Series Precisions electronic balance
Electronics Equipment Co., Ltd);Single clean work station (SW-CJ-1D);Shaken cultivation case (HZQ-F160 types) (Harbin City east
Connection electronic technology development corporation, Ltd.);TG-16-W high speed centrifugal machine for minim;Polarizing microscope (Nikon CHINA YS2-
H1112438);Transmission electron microscope JEM-2100 (Jeol Ltd.);Optical microscope (NO.970810);PH is counted
(sartorius PB-10);X-ray diffraction (XRD) (2036E202 of Ultima IV);Atomic absorption spectrophotometer TAS986
(Beijing Pu Xi general finites company);Ultrasonic washing instrument (KQ-250B);Fourier transform infrared spectrometer
(Nicolet380);Electro-heating standing-temperature cultivator (Kunshan Ultrasonic Instruments Co., Ltd.).
4. culture medium
(1) SRB2 liquid seeds
The Carnis Bovis seu Bubali cream tryptone liquid culture based formulas of table 1
PH to 7.2 is adjusted with the sodium hydroxide solution of 1mol/L, then 121 DEG C of high temperature sterilize 20min.
(2) SRB2 solids seed culture medium
The beef extract-peptone solid culture based formulas of table 2
PH to 7.2 is adjusted to add agar with the sodium hydroxide solution of 1mol/L, then 121 DEG C of high temperature sterilize 20min.
(3) cytoalgae culture medium
The configuration of BG-11 culture medium:
1) citric acid 0.3g, ferric ammonium citrate 0.05g, plus distilled water to be settled to 100mL standby;
2) sodium nitrate 30g, dipotassium hydrogen phosphate 0.78g, bitter salt 1.5g.Distilled water is added to be settled to 1000mL standby
With;
3) sodium carbonate 2g, adds distilled water to be settled to 100mL standby;
4) CALCIUM CHLORIDE DIHYDRATE 1.9g, adds distilled water to be settled to 100mL standby;
5)A5The configuration of micro solution:Boric acid 2.86g, four chloride hydrate manganese 1.81g, Zinc vitriol 0.222g, two
Molybdic acid hydrate sodium 0.39g, copper sulphate pentahydrate 0.079g, cabaltous nitrate hexahydrate 0.0494g.Plus distilled water is settled to 1000mL.4℃
Save backup;
Take 2,20,2,1,1mL from the solution for 1), 2), 3), 4), 5) having configured respectively to be added in 1000mL volumetric flasks,
It is subsequently adding distilled water constant volume.
(2) regulation of culture fluid pH:PH to more than 7.2 is adjusted with the NaOH of 1mol/L;Subpackage is carried out using conical flask to go out
Bacterium.
(3) inoculation of cytoalgae and culture:The OD for measuring mother solution is 673nm, and cytoalgae is accessed on superclean bench, is pressed
According to per bottle 1% of inoculum concentration (adding 2mL per 200mL), move into afterwards in illumination box.Intensity of illumination is 5000lx, and temperature is
25 DEG C, illumination in 12 hours, dark continuous culture in 12 hours.
(4) inducing culture
.Ca in experimentation2+、Mg2+Concentration proportioning is accurately weighing CaCl2·2H2O、MgCl2·6H2The shape of O solids
Formula is obtained.Due to and CaCl2·2H2O and MgCl2·6H2The aerial water absorption of O is very strong, so needing when preparing rapidly to subtract
Few moisture absorption.By CaCl2Solution concentration is set to 1.0mol/L, and further according to sample volume corresponding amount is added, you can obtain initial
Calcium ion concentration is the precipitation environment of l0.0mmol/L.In the same manner by solution MgCl2·6H2O concentration is set to 2.0mol/L, then root
Corresponding amount is added according to sample volume, you can obtain the precipitation environment that initial magnesium ion concentration is respectively 60.0mmol/L.
Using CaCl2·2H2O prepares 100mL1.0mol/LCa2+Solution, needs the CaCl for accurately weighing 14.7g2·
2H2O, in being dissolved in a small amount of distilled water, whisks dissolving, until being completely dissolved and solution change clarification.Then by the solution in beaker
In being transferred to 100mL volumetric flasks, with distilled water rinse 3 times, it is ensured that solute is transferred completely in volumetric flask rear constant volume.
Using the magnesium ion solution for preparing the above-mentioned concentration of l00mL, the MgCl of 40.6g need to be weighed2·6H20, it is dissolved in a small amount of
In distilled water, dissolving is whisked, until being completely dissolved and solution change clarification.Then the solution in beaker is transferred to into 100mL capacity
In bottle, with distilled water rinse 3 times, it is ensured that solute is transferred completely in volumetric flask rear constant volume.
(5) Na needed for precipitating2CO3And NaHCO3Solution
Accurately weigh the sodium carbonate solid of 10.6g, in being dissolved in 50mL distilled water, stirring and dissolving, until be completely dissolved and
Solution becomes clarification.8.4g sodium bicarbonate solids are accurately weighed, in being dissolved in a small amount of distilled water, 100mL volumetric flasks is then transferred to
Middle addition distilled water constant volume is standby.
5. experimental technique
(1) preparation of Bacillus licheniformis
Bacillus licheniformis (Bacillus licheniformis SRB2) are taken from the deep freezer of -4 DEG C of preservation
Go out, configure 5 different dilution gradients, the diluent for taking 800 μ L of each gradient is coated on flat board corresponding to each gradient
On, each gradient does three Duplicate Samples.Flat board is placed in 28 DEG C of incubator, is cultivated 1-2 days.Take growing way preferably single bacterium
Fall, be inoculated in 2 250ml conical flasks for filling 100ml aseptic seed fluid mediums with the ring of inoculating loop picking 2, then put
Shaken cultivation 1-2 days in 28 DEG C of shaken cultivation casees, rotating speed is 130r/min.
(2) preparation of cytoalgae
The seed liquor of cytoalgae PCC8603 is inoculated into the BG- for preparing according to the inoculum concentration of 1% (meeting 1mL per 100mL)
In 11 culture medium, move into afterwards in illumination box.Intensity of illumination is 5000lx, and temperature is 25 DEG C, illumination in 12 hours, 12 hours
Dark continuous culture.
(3) foundation of bacterium-algae syntaxial system
The Bacillus licheniformis and cytoalgae that are respectively grown to stable phase are carried out into the foundation of syntaxial system, is determined respective
Light absorption value, proportionally 1:50;1:100;1:500;1:1000 and phycomycete ratio 1:100000;1:10000;1:1000;1:
100;1:10;1:1;0:1;0:0 carries out Hybrid NC machine tool, and condition of culture is identical with the condition of culture of cytoalgae.
(4) growth curve
The growth curve of microorganism is the curve of cell quantity increase and growth time relation when representing that microbial body grows.
With the antibacterial that fragmentation mode is bred, after being inoculated in fluid medium, under suitable growth conditionss, with bacterial cell number
Logarithm is vertical coordinate, and the growth curve that growth time is plotted by abscissa can be divided into four major parts, reflects antibacterial life
Four long Main Stages:Period of delay, exponential phase, resting stage and decline phase.
Because the concentration of bacterial suspension is directly proportional to absorbance (OD values), therefore using spectrophotometric determination bacteria suspension
Absorbance deducing the concentration i.e. bacterial density of bacterium solution.Measure Bacillus licheniformis, the growth curve of cytoalgae is for side
Just determine that incubation time is respectively to Bacillus licheniformis, the impact of cytoalgae growth in experiment, while measuring Bacillus licheniformis
With cytoalgae coupled growth inducing culture under the conditions of Mg/Ca ratios OD600、OD673Over time situation, is for convenience
Speculate that inducing culture induces the situation of carbonate mineral formation.
The growth tendency of Bacillus licheniformis:Respectively in 0h, 3h, 6h, 9h, 12h, 15h, 24h, 29h, 34h, 39h, 49h
When, the culture fluid of the Bacillus licheniformis in each conical flask is taken, with UNIC722S type ultraviolet-uisible spectrophotometers with right
In the case that the culture medium answered makees blank, absorbance of the bacterium solution under 600nm wavelength is measured, with thunder magnetic pHS-3C type pH meters
The pH value of measurement bacterium solution, measures three parallel laboratory test groups, the OD averaged as this600, pH measured value.So survey altogether
Amount sample 10 times, obtains 10 groups of relevant data, then with the time as abscissa, respectively with OD600It is vertical coordinate with pH, draws out
The OD of Bacillus licheniformis600, pH situations over time.
The growth tendency of cytoalgae:Sample daily in 1-2 days after cytoalgae culture, every 2 days in 2-16 days
Sampling, sampled in 16-40 days every 3 days, sampled every 5 days in 40-65 days, used spectrophotometric determination OD673Value,
The pH value of bacterium solution is measured with thunder magnetic pHS-3C types pH meter.So be total to measuring samples 23 times, obtain 23 groups of relevant data, then with when
Between be abscissa, respectively with OD673, pH be vertical coordinate, draw out growth change situation of the cytoalgae with the time.
The growth tendency of bacterium-algae symbiosis:After bacterium-algae syntaxial system is set up, sampled daily in 1-3 days, 3-
Sampled every 3 days in 11 days, determine Bacillus licheniformis, the OD values of cytoalgae under 600,673 wavelength respectively.So survey altogether
Amount sample 7 times, obtains 7 groups of relevant data, then with the time as abscissa, respectively with OD600、OD673For vertical coordinate, bacterium is drawn out
Algae with the time growth change situation.
(5) atomic absorption spectrophotometer measurement Ca2+Concentration change
Na is added after medium sterilization2CO3And NaHCO3Solution is followed by before bacterium just sampling, is designated as the 0th day, hereafter every
Sampling in one day once, is sampled 7 times altogether.Sampling method:Culture fluid 3mL is taken every time in the centrifuge tube of 5mL, and Jing is centrifuged (10000r/
Min, 5min) take the used membrane filtration of supernatant and obtain filtrate 1mL in the centrifuge tube of 10mL, then add 9mL in centrifuge tube
Distilled water, i.e., bacterium solution dilute 10 times.The purpose of dilution is to prevent atomic absorption spectrophotometer from being blocked by bacterium solution, while also can
Reduce and each take liquid measure not affect the normal growth of thalline.
Calcium ion standard solution is configured:Configuration calcium ion mother solution, takes anhydrous calcium chloride, 150 in electric drying oven with forced convection
DEG C drying three hours, weigh calcium chloride 1.3875g, be settled to 1L, now calcium ion concentration be 500mg/L, 1mL is taken respectively,
2mL, 3mL, 4mL, 5mL, 6mL are added to constant volume in 100mL volumetric flasks, and calcium ion concentration is respectively 5mg/L, 10mg/L,
15mg/L, 20mg/L, 25mg/L and 30mg/L.Take respectively standby in 10 milliliters of immigration centrifuge tubes.Take 10 milliliters of distilled water to add
In centrifuge tube, its calcium ion concentration is 0mg/L.A flame atomic absorption spectrophotometer is often used, a calcium ion need to be matched somebody with somebody
Titer.
Determining Ca by Atomic Absorption Spectrometry ion concentration:First light absorption value is determined with calcium ion titer, with calcium ion concentration as horizontal seat
Mark, light absorption value makes standard curve for vertical coordinate.Afterwards the absorbance of determination sample, show that calcium ion is dense according to standard curve
Degree.
(6) polarized light microscope observing
Sampling in the 56th day after bacterium-algae syntaxial system is set up carries out polarized light microscope observing.Sampling method:Take culture
Basilar parts are deposited in 1.5mL centrifuge tubes, are first stood 10min and are discarded a small amount of liquid of the top with a large amount of thalline, then Jing centrifugations
(10000r/min, 3min) abandoning supernatant, then each centrifuge tube respectively add 1mL distilled water in centrifuge tube, it is vibration, mixed
It is even, then Jing centrifugation (10000r/min, 3min) abandoning supernatants, so with three precipitations of distilled water wash.It is eventually adding about
The dehydrated alcohol of 1mL, after vibration, mixing, takes a small amount of drop that mixes on microscope slide, is placed under micropolariscope and observes.For
Apparent observation mineral surfaces pattern, amplification is 200 times.
(7) Fourier transformation infrared spectrometer analysis
Sampling in the 56th day after bacterium-algae syntaxial system is set up carries out infrared spectrum analysiss in Fu.Sampling method:Take culture
Basilar parts are deposited in 1.5mL centrifuge tubes, are first stood 10min and are discarded a small amount of liquid of the top with a large amount of thalline, then Jing centrifugations
(10000r/min, 3min) abandoning supernatant, then each centrifuge tube respectively add 1mL distilled water in centrifuge tube, it is vibration, mixed
It is even, then Jing centrifugation (10000r/min, 3min) abandoning supernatants, so with three precipitations of distilled water wash.It is eventually adding about
The dehydrated alcohol of 1mL, vibration, mix after stand, bottom precipitation is taken on microscope slide, with slide by pressed powder after natural drying
Rear and potassium bromide is scraped according to 1:100 mixing, after being fully ground.
(8) XRD analysis
Samplings in the 20th, 35,56 days after bacterium-algae syntaxial system is set up carry out XRD analysis.Sampling method:Take culture medium
Bottom precipitation stands 10min in 10mL centrifuge tubes, first and discards medium liquid of the top with a large amount of thalline, then bottom is sunk
Shallow lake is proceeded in 1.5mL centrifuge tubes, Jing centrifugation (10000r/min, 3min) abandoning supernatants, and then each centrifuge tube respectively adds 1ml
Distilled water, with the precipitation of distilled water wash three times, remove salt ion.It is eventually adding quiet after the ethanol of about 3 times of precipitation volumes
Put, with liquid-transfering gun ethanol is sucked, take bottom precipitation and XRD is on coverslip.XRD scanning angles are 10 ° -90 °, and step-length is
0.02,8 °/min of scanning speed.
6. result and analysis
(1) measure of growth curve
1) measure of Bacillus licheniformis growth curve
Because the concentration of bacterial suspension is directly proportional to absorbance (OD values), therefore using spectrophotometric determination bacteria suspension
Absorbance deducing the concentration i.e. bacterial density of bacterium solution.Respectively 0h, 3h, 6h, 9h, 12h, 15h, 24h, 29h, 34h, 39h,
Sample during 49h, with UNIC722S types ultraviolet-uisible spectrophotometer in the case where blank is made with corresponding culture medium, survey
Absorbance of the amount bacterium solution under 600nm wavelength.With the time as abscissa, with OD values as vertical coordinate, Bacillus licheniformis are drawn out
Growth curve, as shown in Figure 1, growth of the Bacillus licheniformis in beef-protein medium includes the figure for obtaining
Three continuous stages:(1) lag phase, now growth conditionss of the antibacterial just in acclimatizing culture medium, curve can from figure
It is 0-3h to go out, as the lag phase of Bacillus licheniformis growth.(2) logarithmic (log) phase, represents fissional active state, from figure
In it is seen that 3-36h, i.e., interior Bacillus licheniformis mushroom out during this.(3) stable phase, now due in culture medium
Nutrient substance be consumed and toxic metabolic products gradually accumulation, the growth rate of antibacterial slows down gradually but total biomass
Still remain unchanged, and the growth rate of Bacillus licheniformis is suitable with decline decomposition rate, thus bacterium solution total biomass is constant,
Show as bacterium solution absorbance constant.It can be seen that being stable phase after 36h, bacterium solution luminosity rate of rise is zero.
2) cytoalgae growth curve is determined
The seed liquor of cytoalgae PCC8603 is inoculated into the BG- for preparing according to the inoculum concentration of 1% (meeting 1mL per 100mL)
In 11 culture medium, move in illumination box and cultivate.Intensity of illumination is 5000lx, and temperature is 25 DEG C, illumination in 12 hours, 12 hours
Dark continuous culture.With the time as abscissa, with OD values as vertical coordinate, using the suction of spectrophotometric determination cytoalgae bacteria suspension
Luminosity is the density of cytoalgae come the concentration that deduces bacterium solution, draws out the growth curve of cytoalgae, the figure for obtaining such as accompanying drawing 2
Shown, growth of the cytoalgae in BG-11 culture medium includes three continuous stages:(1) lag phase, now cytoalgae fit
Answer the growth conditionss in culture medium, the curve from figure it is seen that 1-2 days, the as lag phase of cytoalgae growth.(2) logarithm
Phase, fissional active state is represent, be as can be seen from the figure 2-50 days, i.e., interior cytoalgae is mushroomed out during this.
(3) stable phase, now because the nutrient substance in culture medium is consumed and the gradually accumulation of toxic metabolic products, cytoalgae
Growth rate slows down gradually but total biomass still remains unchanged, and the growth rate of cytoalgae and decline decomposition rate phase
When, thus total biomass is constant in culture fluid, that is, show as culture fluid absorbance constant.It can be seen that after 50 days
As stable phase, culture fluid absorbance rate of rise is zero.
3) growth curve for mixing Bacillus licheniformis in thalline system is determined
After bacterium-algae syntaxial system is set up, sampled daily in 1-3 days, sampled every 3 days in 3-11 days, 600
Wavelength under determine Bacillus licheniformis OD values.So measuring samples 7 times altogether, obtain 7 groups of relevant data.With the time as horizontal seat
Mark, with OD600For vertical coordinate, the growth change curve for drawing out the OD values of Bacillus licheniformis in syntaxial system with the time is for example attached
Shown in Fig. 3, after setting up bacterium-algae syntaxial system, on 1-2 days in syntaxial system of the cell quantity of Bacillus licheniformis are quick
Rise;In 2-7 days, the growth pattern of cell quantity tends towards stability;Growth is presented always afterwards ascendant trend in 7 days.Thus draw:
Bacillus licheniformis and cytoalgae can be set up altogether using the physiological function synergism between algae and the class of antibacterial two biology
Raw body system, and setting up syntaxial system seven days afterwards, Bacillus licheniformis and cytoalgae ratio are 1:Thalline in 100 system
Growth is most fast, peak value occurs.
4) growth curve for mixing cytoalgae in thalline system is determined
After bacterium-algae syntaxial system is set up, sampled daily in 1-3 days, sampled every 3 days in 3-11 days, 673
Wavelength under determine cytoalgae OD values.So measuring samples 7 times altogether, obtain 7 groups of relevant data.With the time as abscissa, point
Not with OD673For vertical coordinate, draw out the OD values of cytoalgae in syntaxial system with the time growth change curve as shown in Figure 4,
After setting up bacterium-algae syntaxial system, Synechocystis cell quantity presenting for 1-2 days in syntaxial system rises;In 2-5 days, cell
Quantity is presented and declined;Growth is presented always afterwards ascendant trend in 5 days.Thus draw:Growth of the Bacillus licheniformis to cytoalgae
Have an impact, set up second day of syntaxial system and the transformation of growth tendency occur within the 5th day;Bacillus licheniformis and cytoalgae
Ratio is 1:50 at second day there is peak value.
(2) ensaying of bacterium-algae symbiosis induction
1) the culture medium bottom precipitation for having cultivated the different bacterium algae ratios of the Mg/Ca=6 of a period of time is analyzed, is tied
Fruit bacterium algae ratio is respectively:1:1000;1:500;1:100;1:50, it can be deduced that:Bacterium-algae syntaxial system can be with after setting up
Ore deposit is induced into, with the prolongation of incubation time, the mineral of induction increase therewith;The product ore deposit number of the syntaxial system of different bacterium algae ratios
Amount is different, and 1:500 and 1:Ore deposit is produced in 100 system more.
2) atomic absorption spectrophotometer measurement Ca2+Concentration change
This measuring Ca2+The sampling mode of concentration change is:Sample after inoculation once, one is every other day sampled afterwards
It is secondary, sample 7 times altogether, measure Ca with atomic absorption spectrophotometer2+Concentration change, makees curve such as accompanying drawing 5, and wherein a is control
Group Ca2+Concentration change figure, b is experimental group Ca2+Concentration change figure, it can be deduced that:Second day after inoculation, matched group and reality
Test Ca in group2+Concentration is remarkably decreased, and this is the CO due to adding when culture medium is prepared3 2-And HCO3 -With the Ca in culture medium2+
A large amount of combination generates precipitation of calcium carbonate.In experimental group during Mg/Ca=6, the mineral of generation are mainly calcite, aragonite, consume
A large amount of Ca2+.The Ca of matched group and experimental group in figure2+Concentration change trend is consistent, but the Ca in matched group2+Concentration all the time will
More than the Ca in experimental group2+Concentration, this effect for being possibly due to microorganism changes the species and pattern of carbonate.
3) polarized microscope analysis of mineral
The mineral in the bacterium-algae syntaxial system cultivated 56 days are taken, polarized light microscopy Microscopic observation (amplifying 200 times) is placed in,
Phycomycete ratio is respectively 1:100000;1:10000;1:1000;1:100;1:10;1:1;0:1;0:0, it can be deduced that:Cultivate
The mineral induced in the different bacterium-algae syntaxial system of 56 days mainly have bar-shaped, spherical, dumbbell shaped, two it is spherical to together
Assembly.Bacterium algae ratio is 1:Mineral in 100000 syntaxial system are predominantly bar-shaped, in addition also dumbbell shaped and two
It is spherical to assembly together, it is a small amount of spherical;Bacterium algae ratio is 1:Mineral in 10000 syntaxial system with bar-shaped in the majority,
In addition also spherical, rhombus, dumbbell shaped and two are spherical to assembly together;Bacterium algae ratio is 1:1000 syntaxial system
In mineral it is predominantly bar-shaped, spherical, also a small amount of two are spherical to assembly together;Bacterium algae ratio is 1:100 are total to
Mineral in raw body system are predominantly bar-shaped, two it is spherical to assembly together, also have in addition a small amount of spherical;Bacterium algae ratio is
1:Mineral in 10 syntaxial system are predominantly spherical, in addition also it is a small amount of it is bar-shaped, two it is spherical to assembly together;
Bacterium algae ratio is 1:Mineral in 1 syntaxial system with spherical in the majority, have in addition two it is spherical to assembly together;Bacterium algae
Ratio is 0:Mineral in 1 syntaxial system have spherical, bar-shaped and dumbbell shaped assembly based on dumbbell shaped, in addition;Bacterium algae
Ratio is 0:Mineral in 0 syntaxial system have in addition two spherical and multiple spherical groups got together with spherical in the majority
It is fit;
According to bacterium, the respective growing state of algae in bacterium-algae syntaxial system, so as to increase Bacillus licheniformis in homobium
Ratio in system is cultivated again.The mineral in bacterium-algae syntaxial system that this time has been cultivated 28 days are taken, polarized light microscopy is placed in
Microscopic observation (amplifies 200 times), and bacterium algae ratio is respectively:1:1000,1:500,1:100,1:50, it can be deduced that:Cultivate 28
The mineral that induce in it different bacterium-algae syntaxial system mainly have bar-shaped, spherical, dumbbell shaped, two it is spherical to together
Assembly.Bacterium algae ratio is 1:Mineral in 1000 syntaxial system are predominantly bar-shaped, spherical, also a small amount of two it is spherical to
Assembly together;Bacterium algae ratio 1:Mineral in 500 syntaxial system with spherical in the majority, it is also a small amount of bar-shaped and two it is spherical
To assembly together;Bacterium algae ratio is 1:Mineral in 100 syntaxial system are predominantly bar-shaped, two it is spherical to together
Assembly, also have in addition a small amount of spherical;Bacterium algae ratio is 1:Mineral in 50 syntaxial system are in the majority with spherical, dumbbell shaped,
Also a small amount of is bar-shaped and two or more spherical to assembly together.
To sum up experimental result can draw:Spherical mineral are only existed in all non-existent cultivating system of bacterium algae;Only algae is deposited
Cultivating system in based on dumbbell shaped mineral;Increasing with Bacillus licheniformis in syntaxial system, the ore deposit of other forms
Thing is gradually decreased, and dumbbell shaped and spherical mineral increase.Illustrate that thalline has a certain degree of impact for bacterium-algae symbiosis into ore deposit.
Can draw through consulting literatures:Spherical mineral are modal mineral combination volume morphings in this experiment, especially
In higher Mg/Ca than in sample.It is the final form of carbonate mineral that Buczynski et al. thinks spherical.Therefore can be with
Speculate, ore-forming element content is higher when Mg/Ca is higher, once nucleation, it is substantial amounts of into ore deposit ion aggregation together, in Organic substance
Promotion, adjustment effect under quick form spheroidal minerals aggregation.Another kind of relatively common form is dumbbell in testing herein
Shape.Buczynski et al. (1991) thinks that dumb-bell shape is distinctive mineral shape under bacterial action.Many researchs think, nothing
It is the universal form of biogenesis carbonate mineral (such as containing magnesian calcite, aragonite etc.) by being dumb-bell shape or spherical.It is mute
Bell-shaped mineral aggregate is probably the higher carbonate mineral of magnesium content, it is also possible to Ca2+、Mg2+Under the induction of Organic substance
Crystal both sides are assembled in a large number, while CO3 2-Shift to both sides and Ca2+、Mg2+With reference to, as a result cause both sides to grow and accelerate, middle C axles
Growth becomes relatively slow, forms dumb-bell shape, and then is grown to by dumb-bell shape spherical.
4) Fourier transformation infrared spectrometer analysis
The bacterium algae ratio for taking Mg/Ca=6 is respectively 1:100000、1:10000、1:1000、1:100、1:10、1:1、0:1、
0:The mineral in the bacterium-algae syntaxial system of 56 days are cultivated under conditions of 0, Fourier transform infrared spectrometer analysis has been carried out, has been obtained
The result for arriving such as accompanying drawing 6, searching related data can obtain:
The infrared absorption peaks of the different crystal system calcium carbonate of table 3
Can draw:Find compared with the data according to table 3, cultivated the bacterium-algae homobium of the different proportion of 56 days
The mineral of induced synthesis have calcite, monohydrocalcite, hydromagnesite under the conditions of system.Bacterium algae ratio is 1:100000、1:
10000、1:The mineral of 1000 syntaxial system induction are calcite;Bacterium algae ratio is 1:The ore deposit of 100 syntaxial system induction
Thing is calcite and monohydrocalcite;Bacterium algae ratio is 1:The mineral of 10 syntaxial system induction are calcite and hydromagnesite;
Bacterium algae ratio is 1:The mineral of 1 syntaxial system induction are calcite and monohydrocalcite;Bacterium algae ratio is 0:1 syntaxial system
The mineral of induction are calcite;The mineral for neither connecing bacterium nor connecing the blank control group induction of algae are calcite.Experimental group with it is right
Comparing according to group to find, formation of the bacterium-algae syntaxial system to mineral has certain impact so as to can be formed except side
Mineral beyond Xie Shi.
With reference to the accompanying drawings the bacterium algae ratio of Mg/Ca=6 is respectively 1 in 6:100000、1:10000、1:1000、1:100、1:
10、1:1、0:1、0:The infared spectrum analysis result of the bacterium-algae syntaxial system Minerals of 56 days is cultivated under conditions of 0, has been true
The ratio of fixed more preferably bacterium-algae symbiosis Metallogenic system, reduces bacterium algae proportion, is changed to 1:1000、1:500、1:100、
1:50, then taking experimental group and matched group in the bacterium-algae syntaxial system for cultivated 20,28 days under Mg/Ca=6 is carried out in Fu
Leaf transformation infrared spectrometer is analyzed, the result for obtaining such as accompanying drawing 7, it can be deduced that:B schemes, and c figures are that bacterium algae ratio is respectively 1:
1000、1:500、1:100、1:Cultivated under 50 symbiosis conditions 20 days and 28 days induced synthesis mineral infrared spectrum.With
Data in table 3 compare discovery, and the mineral of induced synthesis are mainly calcite, aragonite under the conditions of syntaxial system;Neither connect bacterium
The mineral for not connecing the blank control group induction of algae are calcite.Identical with accompanying drawing 6, experimental group can find compared with matched group,
There is certain impact in formation of the bacterium-algae syntaxial system to mineral.The time of syntaxial system induction mineral makes a farfetched comparison Fig. 6 in accompanying drawing 7
Time it is short, do not formed except calcite, other mineral beyond aragonite.Continue to extend such as incubation time, meeting
There are other mineral.
Consulting literatures are learnt:Hydrone has stronger quiet in the presence of hydrogen bond as polar molecule to the ion in solution
Electric attraction.Mg and Ca respectively positioned at the second main group the 3rd, the period 4, the atomic radius of the atomic radius of Ca than Mg is big,
So Mg2+Easily form hydrate with water.Therefore, Mg is difficult to be combined by carbonate in natural environment, it is more difficult to enter into
In Calcium Carbonate lattice, and easily combined with water.It can be said that Mg2+Hydration hinder magnesium ion in solution and enter carbonic acid
Calcium crystal grid.
6 understand to define hydromagnesite in bacterium-algae syntaxial system with reference to the accompanying drawings, it may be possible to because the bar existed in bacterium algae
Magnesium ion in solution is set to be more easy to be combined to induce the formation of hydromagnesite with carbonate under part.
5) XRD analysis
According to the observation situation of mineral under polarizing microscope under conditions of identical Mg/Ca=6 in accompanying drawing 7, in order to ore deposit
Thing is qualitatively analyzed, and confirms the species of mineral, and then has carried out the experiment of mineral X-ray diffraction analysis (XRD).X-ray
During by crystal, peak spectrogram as shown in Figure 8 can be produced.The corresponding interplanar distance of each spectral peak has with shapes and size
Close, the relative intensity of peak value is relevant with the species of crystal particle and position, also related to degree of crystallinity, the relatively low spectral peak of relative intensity
Mean that the crystallization degree of the mineral is weaker, and the high spectral peak of relative intensity then shows that crystallization degree is higher.Take and cultivated 20,
35th, the experimental group and matched group in the bacterium of 56 days-algae syntaxial system under Mg/Ca=6 is XRD, the result for obtaining such as accompanying drawing 8, its
In, a, b, c are respectively the XRD spectrum for having cultivated 20,35,56 days mineral, it can be found that:
(1) mineral for having cultivated the experimental group of 20 days are mainly calcite.Bacterium algae ratio is respectively 1:100000、1:
10000、1:1000、1:100、1:10、1:1、0:The mineral induced under the conditions of 1 are calcite;Neither connect bacterium nor connect algae
The mineral that induce of matched group be calcite and aragonite.
(2) mineral for having cultivated the experimental group of 35 days are mainly calcite.Bacterium algae ratio is 1:100000、1:10000、1:
1000、1:100、1:10、1:1、0:The mineral induced under the conditions of 1 are calcite.Neither connect bacterium nor connect the matched group of algae
The mineral for inducing are calcite and aragonite.
(3) mineral for having cultivated the experimental group of 56 days are mainly calcite, also Du Pingshi, hydromagnesite, aragonite, Dan Shui
Calcite.Bacterium algae ratio is respectively 1:100000、1:10000、1:The mineral that 1000 system is induced are mainly calcite, Du
Flat stone.Bacterium algae ratio is 1:The mineral that 100 system is induced are mainly calcite, monohydrocalcite, Du Pingshi.Bacterium algae ratio
For 1:The mineral that 10 system is induced are mainly calcite, hydromagnesite.Bacterium algae ratio is 1:The mineral that 1 system is induced
Predominantly calcite, monohydrocalcite.Bacterium algae ratio is 0:The mineral that 1 system is induced are calcite, aragonite, hydromagnesite;
It is calcite and aragonite neither to connect bacterium nor connect the mineral that the matched group of algae induces.
Can draw:Bacterium-algae syntaxial system can induce into ore deposit, and as the prolongation of incubation time engenders
Other mineral in addition to calcite is present, including Du Pingshi, hydromagnesite, aragonite, monohydrocalcite.The ratio of bacterium algae is different
The mineral for being formed also difference, bacterium algae ratio is respectively 1:100000、1:10000、1:The ore deposit formed under conditions of 1000
Thing is similar, illustrates the dense impact very little to mineral of bacterium in this proportion.Find through contrast, with the raising that bacterium is dense,
The increase of bacterium algae ratio, the absworption peak of mineral not only increases but also becomes strong.Illustrate the dense higher training to bacterium-algae syntaxial system of bacterium
It is foster to affect bigger, be more conducive to into ore deposit.
With reference to the accompanying drawings the bacterium algae ratio of Mg/Ca=6 is respectively 1 in 8:100000、1:10000、1:1000、1:100、1:
10、1:1、0:The XRD spectrum analysis result of the bacterium-algae syntaxial system Minerals of 20,35,56 days is cultivated under conditions of 1, has been
It is determined that the more preferably ratio of bacterium-algae symbiosis Metallogenic system, reduces bacterium algae proportion, 1 is changed to:1000、1:500、1:
100、1:50, then take experimental group and matched group in the bacterium-algae syntaxial system for cultivated 20,28 days under Mg/Ca=6 and do
XRD, the wherein result for obtaining such as accompanying drawing 9, d, e are respectively the XRD spectrum for having cultivated 20,28 days mineral.
It can be found that:Bacterium algae ratio is respectively 1:1000、1:500、1:100、1:The mineral induced under the conditions of 50 are
Calcite and aragonite.It is also calcite and aragonite neither to connect bacterium nor connect the mineral that the matched group of algae induces.
Can draw:The mineral for having cultivated the experimental group of 20,28 days are mainly calcite and aragonite.In accompanying drawing 9 d figure with
E figures compare discovery, and the absworption peak in e figures is significantly more than d figures, and intensity is high.Illustrate with the prolongation of incubation time, culture
Mineral in system there occurs change, and difference occurs in the peak spectrogram for causing XRD.The a figures that the d figures of accompanying drawing 9 are compared to accompanying drawing 8 are equal
To have cultivated the mineral peak spectrogram of 20 days, the mineral for being derived have also appeared aragonite except calcite.But it is observed that can be with
It was found that, it is both in the syntaxial system of different bacterium algae ratios and has cultivated the peak height that the mineral of 20 days are showed, peak separation, peak intensity
Degree is different.The intensity for being vaccinated with the mineral characteristic peak of the cultivating system induction of different bacterium algae ratios is significantly stronger than and neither connect bacterium
The blank control group of algae is not connect, illustrates that microorganism uprises can the crystallization degree of carbonate mineral yet.The peak spectrogram chats of accompanying drawing 9
The induction time of thing makes a farfetched comparison the short of Fig. 8, does not occur that, except calcite, the diffraction maximum of other mineral of aragonite, one may be further cultured for
Can change after the section time.
Experiment conclusion:
By experimental verification, after Bacillus licheniformis are mixed with cytoalgae, the cell quantity of Bacillus licheniformis exists
1-2 days rapid increases in syntaxial system;In 2-7 days, the growth pattern of cell quantity tends towards stability;7 days latter of growth
It is straight that ascendant trend is presented.Thus draw:Bacillus licheniformis and cytoalgae can be using between algae and the class of antibacterial two biologies
Physiological function synergism establishes syntaxial system, and setting up syntaxial system seven days afterwards, Bacillus licheniformis and collection born of the same parents
Algae ratio is 1:Thalli growth is most fast in 100 system, peak value occurs.
Set up Synechocystis cell quantity presenting for 1-2 days in syntaxial system after syntaxial system to rise;In 2-5 days, carefully
Born of the same parents' quantity is presented and declined;Growth is presented always afterwards ascendant trend in 5 days.Thus draw:Life of the Bacillus licheniformis to cytoalgae
Length has an impact, and set up second day of syntaxial system and the transformation of growth tendency occurs within the 5th day;Bacillus licheniformis and cytoalgae
Ratio be 1:50 at second day there is peak value.
By micropolariscope it is observed that the mineral formed in the experimental group for connecing bacterium algae and the matched group for not connecing bacterium
Form is had any different, and in all non-existent cultivating system of phycomycete spherical mineral are only existed;With mute in the cultivating system that only algae is present
Based on bell-shaped mineral;Increasing with Bacillus licheniformis in syntaxial system, the mineral of other forms are gradually decreased, dumbbell shaped and
Spherical mineral increase.Illustrate that thalline has a certain degree of impact for bacterium-algae symbiosis into ore deposit, microorganism can pass through itself
Metabolic activity change mineral pattern.
By carrying out FTIR spectrum analysis to the solid mineral composition in matched group and experimental group and XRD analysis can
To find:The intensity at the mineral characteristic peak induced in the experimental group of bacterium-algae symbiosis is apparently higher than neither connecing bacterium nor connect the sky of algae
White matched group, illustrates that microorganism can make the aspects such as the crystal grain of carbonate mineral, crystal face more preferably reflect crystallization degree optimization;No
The mineral induced in the syntaxial system of same bacterium algae ratio are different, illustrate that micro organism quantity can change the kind of carbonate
Class.
Claims (10)
1. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system, it is characterised in that:Including what is be sequentially communicated from top to bottom
Bactericidal unit, microorganism reactor, mineral recovery apparatus, are provided with cytoalgae and bacillus cereuss in the microorganism reactor.
2. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 1, it is characterised in that:It is described
Microorganism reactor includes the housing (1) of transparent configuration, and on the side wall of the housing (1) illuminator, the cytoalgae are provided with
It is scattered in the liquid phase in housing (1), the vertical reaction column (3) for being provided with tubular, the reaction column (3) in the housing (1)
Inner chamber in filled with embedding bacillus cereuss sodium alga acid capsule (4), the bottom of the reaction chamber is provided with aerator
(5)。
3. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 2, it is characterised in that:It is described
Device (2) is communicated with the side wall of housing (1), the linker (2) is transparent configuration.
4. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 2, it is characterised in that:It is described
Housing (1) for lucite structure cylindrical housings (1), the reaction column (3) is lucite cylinder structure, the light source
Lamp is LED.
5. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 2, it is characterised in that:It is described
Aerator (5) includes the air inlet pipe (6) for being filled with sterilizing gas, and the outer end of the air inlet pipe (6) is provided with air intake valve, institute
State air inlet pipe (6) and be provided with coupled logical aeration tube (7), at the top of the aeration tube (7) solarization air cap (8) is provided with.
6. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 1, it is characterised in that:It is described
Bacillus cereuss are Bacillus licheniformis.
7. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 1, it is characterised in that:It is described
Bactericidal unit includes being provided with the asbestos heating plate for high temperature sterilize in sterilization tank (9), the sterilization tank (9), tells sterilizing
Inlet (10) is provided with the top of case (9), the sterilization tank (9) is provided with flow control valve and microorganism reactor between.
8. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 7, it is characterised in that:It is described
The layering dividing plate (11) of several levels is provided with sterilization tank (9), layering dividing plate (11) is relative with the section of sterilization tank (9)
Should, one end of the sterilization tank (9) is provided with breach (19), and the breach (19) of adjacent layering dividing plate (11) is shifted to install, institute
State layering dividing plate (11) and be provided with several vertical baffle plates (12), the baffle plate (12) is crisscross arranged to form serpentine channel.
9. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 8, it is characterised in that:It is described
Layering dividing plate (11) is 2, and the baffle plate (12) is 2.
10. the sewage-treatment plant of cytoalgae-bacillus cereuss co-mixing system according to claim 1, it is characterised in that:It is described
Mineral recovery apparatus include reclaiming bucket (13), and described recovery be provided with the discharging being connected with microorganism reactor at the top of bucket (13)
Passage (14), is provided with discharge gate on the discharge channel (14), the side wall top of the recovery bucket (13) is provided with waste liquid row
Mouth (20) is put, the bottom surface for reclaiming bucket (13) is that taper is domatic, and placed in the middle being provided with recovery bucket (13) is rotated with it
The axis (15) of connection, the bottom of the axis (15) is provided with truss (16), and the bottom side of the truss (16) is provided with and reclaims bucket
(13) material scraping plate (17) being engaged, it is described reclaim bucket (13) bottom surface it is placed in the middle be provided with discharging opening (18), the discharging opening
(18) discharge valve is installed on.
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CN106145389A (en) * | 2016-08-25 | 2016-11-23 | 山东科技大学 | The sewage-treatment plant of cytoalgae bacillus cereus co-mixing system and using method thereof |
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