CN103849550A - Secondary culture apparatus and culture method for alga - Google Patents
Secondary culture apparatus and culture method for alga Download PDFInfo
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Abstract
The invention discloses a secondary culture apparatus for alga, which comprises a bottom support plate, a reflective device is fixed on the bottom support plate so as to form a frame, a movable cover plate with holes is carried on the reflective device, and a gas loading device is placed on the movable cover plate with holes; and a light source loading device is carried on the reflective device, and the light source loading device and an alga culture vessel are arranged in the frame. The light source loading device comprises a vertical steel frame, a steel stainless column and a fine column with a fixed ring, wherein the vertical steel frame is arranged at the middle position of the short side edge of the support plate, the steel stainless column is arranged at the middle position of the steel frame, the fine column extends out of the steel stainless column, and a light source is arranged on the steel stainless column; the fixed ring is used for placing the alga culture vessel; the gas loading device comprises a gas pump and a gas inlet pipe connecting the gas pump and the alga culture vessel, and the gas inlet pipe is provided with a gas mass regulating valve and a gas filtering valve; and an opening at the upper part of the culture vessel is provided with a cotton plug or a silica gel plug, and an opening at the bottom of the culture vessel is provided with a liquid outlet valve. The invention also discloses a secondary culture method for alga.
Description
Technical field
The present invention relates to the cultural method of plant plankton, be specifically related to a kind of algae secondary culture apparatus and cultural method thereof.
Background technology
Whole world phycophyta approximately has 40000 kinds, and wherein fresh water algae has 25000 kinds of left and right, and approximately 9000 kinds of the fresh water algaes that China has found.Algae distribution is on earth very wide, has their trace from hot equator to long-term ice-bound polar region.
Algae is not only most important primary producer in aquatic ecosystem, and is the main provider of oxygen in water.Algae produces organic carbon by photosynthesis and fixes nitrogen simultaneously, and the total amount of the organic carbon that algae produces is about 7 times of higher plant, approximately can fix the nitrogen of 1.7 hundred million tons every year.Algae is the direct or indirect bait of fish and other economic animals, significant in the production performance of decision waters.
Algae has advantages of so many, but in the time that " wawter bloom " appears in algae excessive propagation, the harm that production is brought is also difficult to appraisal and even human health is exerted an influence.Take blue-green algae as example, within 2007, Taihu Lake breakout of water bloom causes the difficult gear of Wuxi City tap water stench, cannot drink, and in the serious lake region of situation, lake water is dense thick as green painting, and citizen's life is greatly affected.The same year, Chaohu, Dian Chi were also broken out blue-green algae in succession, and the Important Water Source reservoir in Changchun, North City has also been found blue-green algae.Early 1980s, the Malpas reservoir of large Leah difficult to understand New South Wales, epidemiological study, detects that the poisonous blue-green algae of tap water pollutes the synchronism between liver injury.
The advantage of algae does not also fully demonstrate at present, and the negative impact report of its generation is relatively many, and researchist starts more to pay close attention to this field, and the indoor secondary enlarged culturing of algae is more and more important.Existing patent, mainly for the large scale culturing of algae or the improvement of algae pollution, has no record for the apparatus and method that guarantee scientific research quality.The present invention is convenient to algal grown and is not vulnerable to pollute, and has guaranteed the science of all kinds of researchs, also can observe easily the impact of allogenic material on algal grown simultaneously.Application of the present invention will make the cultivation of algae secondary be convenient to carry out and purity is guaranteed, is convenient to carrying out of multiple research work.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of secondary culture apparatus that guarantees algae normal growth, and this device can make algae stable growth under manual control condition, meets the demand that all kinds of research work are carried out.
The technical problem that the present invention also will solve, is to provide the method for utilizing said apparatus to carry out algae cultivation.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
Algae secondary culture apparatus, comprises bottom support plate, forms framework at the upper fixing reflex reflector of bottom support plate, and the tool hole removable cover carrying on reflex reflector, places gas-loaded device on the removable cover of tool hole; The light source deceleration loading device carrying on reflex reflector is provided with light source deceleration loading device and algae culture vessel in framework.
Wherein, described reflex reflector is that bottom and long side reflector seal, the framework that short side is open,
Wherein, described light source deceleration loading device, for being welded on the vertical steelframe in the short side of back up pad mid-way, steelframe interposition installs several stainless steel columns, extend the buttress shaft with fixed ring from middle stainless steel post, other stainless steel column can need to load varying strength light source according to what cultivate algae kind.Two conventional class fluorescent tubes are growth lamp and LED lamp.
Described gas-loaded device comprises the inlet pipe of air pump and connection air pump and algae culture vessel, and inlet pipe is provided with air volume regulating valve and gas take.
Wherein, described culture vessel comprises tampon or the silica gel plug of culture vessel mouth, air inlet Glass tubing, the liquid outlet of bottom.
Wherein, described tool hole removable cover area covers the opening at reflex reflector top substantially.
Wherein, described reflex reflector is only fixing whole of framework long side, and reflector surface is " Z " shape fold.
Wherein, described light source deceleration loading device, the fixing light source of intermediate column can increase and decrease according to the required light intensity of algal grown, and light source regulates by automatic timing switch.Fixed ring is apart from light source >=30cm, and the upper leaving certain gaps uncovered by the economic plan of circle, is convenient to picking and placeing of algae culture vessel.
Wherein, described culture vessel has scale, tampon tightness have when extracting vessel port " bang " sound be advisable, liquid outlet seals with silicone tube, silicone tube with clip control in case nutrient solution spill.
The invention also discloses a kind of cultural method that utilizes above-mentioned device to commonly use algae, comprise the steps:
A is under 25 ± 2 ℃ of indoor conditionss, and to the nutrient solution that injects aequum in culture vessel, nutrient solution can be tap water, substratum or other water.Culture vessel and water are according to the sterilizing of training objective selectivity.In culture vessel, inoculate algae, concentration reaches 10
4individual/mL;
B culture vessel fixes in algatron, opens light source deceleration loading device, gas-loaded device, observes algae density after moving continuously 4 days.
C, from the culture vessel liquid outlet liquid that takes a morsel, examines under a microscope counting, judges whether to satisfy the demands.
In steps A, described algae initial inoculation concentration need at least reach 10
4individual/mL, pollution-free for the algae inoculating, reach stable logarithmic phase.
In step B, described light source deceleration loading device moved than 16:8 according to the dark time of light.
In step B, described gas-loaded device, adds load volume for forming a string bubble.
Beneficial effect: 1, the present invention is simple and easy to do, workable.Material therefor of the present invention is easy to obtain, simple in structure, manufacture and easy to maintenance.2, the present invention adopt culture vessel once-forming and volume little, avoided the pollution of other material and be easy to change, facilitate succeeding transfer culture; Single culture vessel of algae can load 0～5L solution, and each device can arrange 6, and cultivation amount can meet most of scientific experimentation research.Light intensity deceleration loading device can increase and decrease the different demands of light intensity voluntarily according to cultivating algae kind, and multiple algae can be cultivated simultaneously.3,, after off-test, solution directly after sterilizing, is poured out the workload reducing a staff by liquid outlet.
Accompanying drawing explanation
Fig. 1 is that algal grown of the present invention promotes apparatus structure schematic diagram.
Fig. 2 is light source deceleration loading device structural representation of the present invention.
Fig. 3 is gas-loaded apparatus structure schematic diagram of the present invention.
Fig. 4 is culture vessel structural representation of the present invention.
Embodiment
According to following embodiment, the present invention may be better understood.Those skilled in the art will readily understand, embodiment is only for the present invention is described, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
In the present embodiment, Fig. 1～Fig. 4 Reference numeral is corresponding as follows: 1 tool hole removable cover; 2 reflex reflectors; 3 light source deceleration loading devices; 4 gas-loaded devices; 5 culture vessels; 6 mobile pulleys; 7 light sources; 8 fixed rings; 9 " Z " type reflector; 10 gas takes; 11 air volume regulating valves; 12 inlet pipe; 13 liquid-outlet valves; 14, tampon or silica gel plug; 15, scale; 16, the short side of back up pad; 17, back up pad long side; 18, stainless steel column; 19, buttress shaft; 20, vertical steelframe.
As shown in Figure 1, the present embodiment discloses algae secondary culture apparatus, comprises bottom support plate, forms framework at the upper fixing reflex reflector 2 of bottom support plate, and back up pad is preferentially selected stainless steel; The tool hole removable cover 1 carrying on reflex reflector 2, tool hole removable cover just covers reflex reflector framework; On the removable cover of tool hole, gas-loaded device 4 is set; In framework, be provided with light source deceleration loading device 3 and algae culture vessel 5; Mobile pulley 6 is installed in reflex reflector bottom.Described tool hole removable cover 1 is the fixing synthetic glass of length and width, digs diameter 3cm circular hole and evenly arranges, each 3 of both sides in long edge.
As shown in Figure 2, light source deceleration loading device 3 comprises, be welded on the steelframe 20 in the short side of back up pad 16 mid-ways, steelframe interposition installs several stainless steel columns 18, extend the buttress shaft 19 with fixed ring 8 from two stainless steel columns wherein, other stainless steel column can need to load varying strength light source 7 according to what cultivate algae kind, and light source 7 is by timing device control, according to the dark time of light than 16:8 operation (be to open 16h daytime, close night 8 hours).Fixed ring 8 is apart from light source >=30cm, and the upper leaving certain gaps uncovered by the economic plan of circle, is convenient to picking and placeing of algae culture vessel 5; Reflex reflector is only fixing whole of back up pad long side, and surface is " Z " shape fold, guarantees that light is even.
As shown in Figure 3, gas-loaded device comprises air volume regulating valve 11 and gas take 10.Air volume regulating valve 11 can regulate the gas velocity being charged in culture vessel by knob; 10 liang of side joint silicone tubes of gas take connect air pump and culture vessel, therebetween 0.22um filter membrane.
As shown in Figure 4, culture vessel 4 is the container of the sealing of bottom and sidewall, open-top, and the material of culture vessel 5 is preferably glass, and glass once shaped is without cohesive material, the preferred 0.5cm of wall thickness.Be carved with volume markings outward at culture vessel 5 walls.Inlet pipe 12 passes through to culture vessel 5 bottoms in the middle of tampon 14.
The present embodiment is simple and easy to do, workable, and material therefor is easy to obtain, simple in structure, manufacture and easy to maintenance.The present embodiment adopt culture vessel once-forming and volume little, avoided the pollution of other material and be easy to changing convenient cultivation; Reflex reflector, light source deceleration loading device are beneficial to algae and obtain suitable growth light intensity, accelerating growth.Gas-loaded device and culture vessel are convenient to algae and are obtained sufficient oxygen and be not vulnerable to pollution, have guaranteed the science of all kinds of researchs.The application of the present embodiment will make middle amount algae culture be convenient to carry out and purity is guaranteed.
Embodiment 2:
Utilize above-mentioned device to commonly use a cultural method for algae, the method comprises the steps:
A is under 25 ± 2 ℃ of indoor conditionss, and to the nutrient solution that injects 2L in culture vessel, nutrient solution can be tap water, substratum or other water.Culture vessel and water are according to the sterilizing of training objective selectivity.In culture vessel, inoculate algae, concentration reaches 10
4individual/mL, pollution-free for the algae inoculating, reach stable logarithmic phase.
B culture vessel fixes in algatron, open light source deceleration loading device, gas-loaded device, operation was observed algae density after 4 days continuously, move than 16:8 according to the dark time of light at the light source deceleration loading device described in this process, described gas-loaded device, adds load volume for forming a string bubble.
C, from the culture vessel liquid outlet liquid that takes a morsel, examines under a microscope counting, judges whether to meet work requirements.
Embodiment 3:5 kind is commonly used algal species cultivation
Algae is most important primary producer in aquatic ecosystem, is also the main provider of oxygen in water.Algae is the direct or indirect bait of fish and other economic animals, significant in the production performance of decision waters.Scientific research personnel for algae carry out more and more, indoor secondary enlarged culturing is more and more important.Use secondary enlarged culturing device can conveniently obtain the pure culture liquid of many algaes kind, meet the demand of studying work, this research is cultivated 5 kinds of algae, to verify the upgrowth situation of algae under this device.
1 cultivated material
Chlamydomonas reinhardtii (Chlamydomonas reinhardtii), goat's horn crescent moon algae (Selenastrum capricornutum), Chlorella pyrenoidesa (Chlorella pyrenoidosa), scenedesmus obliquus (Scenedesmus obliquus), chlorella (Chlorella vulgaris Beij.).Algae kind is purchased from hydrobiont institute of Chinese Academy of Sciences algae kind storehouse.
2 culture condition
Plant and instrument: microscope, high-pressure steam sterilizing pan, illumination shaking table, dissolved oxygen meter, thermometer, pH meter, glassware etc.
Envrionment conditions: 23 ± 2 ℃ of temperature, light intensity 5000 ± 500Lx, continuous illumination is cultivated.
Substratum: adopt SE substratum, culture medium prescription is in table 1.
Table 1SE culture medium prescription
Above each composition is mixed with to corresponding mother liquid concentration, and according to indicating that order is transferred to corresponding mother liquor consumption in 1000mL volumetric flask successively, constant volume, through autoclaving (121 ℃, 15min), seals and posts label, 4 ℃ of Refrigerator stores, validity period 2 months.This substratum is used after 10 times of the distilled water dilutings of autoclaving (121 ℃, 15min) and can be used.
* A
5solution formula
EDTA-Fe：
1mol/L HCl: get 4.1mL concentrated hydrochloric acid distilled water diluting to 50mL.
Take 0.1mol/L EDTA-Na
20.9306g is dissolved in 50mL distilled water.
Take FeCl3.6H2O0.901g and be dissolved in the 1mol/L HCl that the above step of 10mL prepared, the 0.1mol/L EDTA-Na2 then having prepared with 10mL mixes, and adds distilled water diluting to 1000mL.
Soil extract: get garden soil and do not execute overfertilization 200g and be placed in beaker or triangular flask, add distilled water 1000mL, bottleneck seals with porous plug, in water-bath, boiling water heating 3h, cooling, precipitation 24h, this process is carried out 3 times continuously, then filter, get supernatant liquor, in high-pressure sterilizing pot, after sterilizing, in 4 ℃ of refrigerators, save backup.
3 algae kind precultures
Test algae kind is equipped with in the triangular flask of SE nutrient solution with sterile manner access, is placed under these conditions in shaking table and cultivates, and transfers once every 4d.Culture transferring 2～3 times, treats frustule in logarithmic phase and reaches synchronous growth, can be used for enlarged culturing.
4 secondary enlarged culturing methods
Get the algae kind 20mL of preculture in logarithmic phase, join in the culture vessel that 2LSE substratum is housed, open light source deceleration loading device and gas deceleration loading device.Regulate light intensity to 4500lux, adjustments of gas deceleration loading device is to the continuous bunchiness bubbling of substratum.In 24h gets culture vessel, algae kind is measured frustule number.
5 cultivation results
The algae density value of measuring each treatment group with blood counting chamber, each nutrient solution at least counts twice, obtains mean value.If twice count results differs larger, answer repeat count.0～72h algal grown measurement result is in table 2.
Average cell number in table 25 class algae kind 72 hours
Test-results shows, test all energy normal growths of 5 class algae kinds used, and cell count rate of growth is good, can practical requirement.
Claims (6)
1. algae secondary culture apparatus, it is characterized in that, comprise bottom support plate, upper fixing reflex reflector (2) at bottom support plate forms framework, at the upper tool hole removable cover (1) carrying of reflex reflector (2), gas-loaded device (4) is set on tool hole removable cover (1), in framework, is provided with light source deceleration loading device (3) and algae culture vessel (5);
Back up pad comprises the short side of back up pad long side (17) back up pad (16), and described reflex reflector (2) is for being fixed on the reflector between back up pad long side (17), and the face of the short side of back up pad (16) opens wide;
Described light source deceleration loading device (3) comprise be arranged on the short side of back up pad (16) mid-way vertical steelframe (20), be arranged on the stainless steel column (18) in steelframe mid-way, and from the buttress shaft (19) of the extended band fixed ring of stainless steel column (8), light source (7) is set on stainless steel column; Described fixed ring is used for placing described algae culture vessel (5);
Described gas-loaded device (4) comprises the inlet pipe (12) of air pump and connection air pump and algae culture vessel (5), and inlet pipe (12) is provided with air volume regulating valve (11) and gas take (10);
Described culture vessel (6) upper opening arranges tampon or silica gel plug (14), and bottom opening arranges liquid-outlet valve (13).
2. algae secondary culture apparatus according to claim 1, is characterized in that, the inner surface of reflector is set to fold face.
3. algae secondary culture apparatus according to claim 1, is characterized in that, described fixed ring (8) apart from light source (7) apart from more than 30cm.
4. utilize device described in claim 1 to commonly use a cultural method for algae, it is characterized in that, comprise the steps:
A, under 25 ± 2 ℃ of indoor conditionss, to injecting nutrient solution in algae culture vessel (5), nutrient solution is tap water or substratum, to inoculation algae in algae culture vessel (5), concentration reaches 10
4more than individual/mL;
B, algae culture vessel (5) fix in fixed ring, open light source deceleration loading device (3), gas-loaded device (4), observe algae density after moving continuously 3～4 days;
C gets about 5ml liquid from algae culture vessel (5) liquid outlet, examines under a microscope counting, judges whether to meet to reach the requirement of logarithmic phase, require index be operation after 4 days algae liquid concentration reach 106/more than mL.
5. the cultural method of conventional algae according to claim 4, is characterized in that, in step B, described light source deceleration loading device (3) moved than 16:8 according to the dark time of light.
6. the cultural method of conventional algae according to claim 5, is characterized in that, described gas-loaded device (4) adds gas carrier requirement and forms a string continuous air bubbles.
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| CN111690540A (en) * | 2020-07-01 | 2020-09-22 | 南县小龙虾协会 | Secondary culture method for algae |
| CN111996103A (en) * | 2020-09-01 | 2020-11-27 | 上海海洋大学 | Multi-mode microalgae culture equipment |
| CN113341078A (en) * | 2021-07-28 | 2021-09-03 | 中国环境科学研究院 | Method and device for rapidly detecting influence on growth of duckweed |
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|---|---|---|---|---|
| CN111690540A (en) * | 2020-07-01 | 2020-09-22 | 南县小龙虾协会 | Secondary culture method for algae |
| CN111996103A (en) * | 2020-09-01 | 2020-11-27 | 上海海洋大学 | Multi-mode microalgae culture equipment |
| CN113341078A (en) * | 2021-07-28 | 2021-09-03 | 中国环境科学研究院 | Method and device for rapidly detecting influence on growth of duckweed |
| CN113341078B (en) * | 2021-07-28 | 2021-12-24 | 中国环境科学研究院 | Method and device for rapidly detecting influence on growth of duckweed |
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| CN103849550B (en) | 2015-09-16 |
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