CN103849550B - A kind of algae secondary culture apparatus and cultural method thereof - Google Patents
A kind of algae secondary culture apparatus and cultural method thereof Download PDFInfo
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Abstract
The invention discloses a kind of algae secondary culture apparatus, comprise bottom support plate, form framework at the upper fixing reflex reflector of bottom support plate, the tool hole removable cover that reflex reflector carries, gas-loaded device placed by tool hole removable cover; The light source deceleration loading device that reflex reflector carries, is provided with light source deceleration loading device and algae culture vessel in framework.Described light source deceleration loading device comprises the vertical steelframe being arranged on back up pad short side mid-way, the stainless steel column being arranged on steelframe mid-way, and the buttress shaft of the band fixed ring extended from stainless steel column, and stainless steel column arranges light source; Described fixed ring is for placing described algae culture vessel; Described gas-loaded device comprises air pump and connects the inlet pipe of air pump and algae culture vessel, and inlet pipe is provided with air volume regulating valve and gas take; Described culture vessel upper opening arranges tampon or silica gel plug, and bottom opening arranges liquid-outlet valve.The invention also discloses algae secondary cultural method.
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 about has 40000 kinds, and wherein fresh water algae has about 25000 kinds, and the fresh water algae about 9000 kinds that China has found.Algae distribution is on earth very wide, has their trace from the equator of sweltering heat to throughout the year 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, about 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 so many advantage, but when " wawter bloom " appears in algae excessive propagation, is also difficult to estimate even has an impact to human health to producing the harm that brings.For blue-green algae, within 2007, Taihu Lake breakout of water bloom causes Wuxi City tap water stench difficulty gear, cannot drink, and in the lake region that situation is serious, lake water is dense thick as green painting, and civil plantation is greatly affected.The same year Chaohu, Dian Chi also breaks out blue-green algae in succession, the Important Water Source reservoir in Changchun, North City have also discovered blue-green algae.Early 1980s, the Malpas reservoir of large Leah New South Wales difficult to understand, 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 that it produces 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.The large scale culturing of existing patent mainly for algae or the improvement of algae pollution, for ensureing that the apparatus and method of scientific research quality have no record.The present invention is convenient to algal grown and is not vulnerable to pollute, and ensure that the science of all kinds of research, also can observe the impact of allogenic material on algal grown easily simultaneously.The cultivation of algae secondary will be made to be convenient to carry out for application of the present invention and purity is ensured, 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 ensureing algae normal growth, and this device can make algae stable growth under manual control condition, meets the demand that all kinds of research work is carried out.
The technical problem that the present invention also will solve, is to provide the method 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, and form framework at the upper fixing reflex reflector of bottom support plate, the tool hole removable cover that reflex reflector carries, gas-loaded device placed by tool hole removable cover; The light source deceleration loading device that reflex reflector carries, 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 back up pad short side mid-way, steelframe interposition installs several stainless steel columns, extend the buttress shaft of band fixed ring from middle stainless steel post, other stainless steel column can according to cultivate algae kind need load varying strength light source.Two conventional class fluorescent tubes are growth lamp and LED.
Described gas-loaded device comprises air pump and connects the inlet pipe of 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 fixed whole of framework long side, and reflector surface is " Z " shape fold.
Wherein, described light source deceleration loading device, the light source that intermediate column is fixed can increase and decrease according to light intensity needed for algal grown, and light source is regulated by automatic timing switch.Fixed ring distance light source >=30cm, and the upper leaving certain gaps uncovered by the economic plan of circle, be convenient to picking and placeing of algae culture vessel.
Wherein, described culture vessel has scale, have when tampon tightness is to extract vessel port " bang " sound be advisable, liquid outlet with silicone tube close, silicone tube clip control in order to avoid nutrient solution spills.
The invention also discloses a kind of cultural method utilizing above-mentioned device to carry out conventional algae, comprise the steps:
A, under 25 ± 2 DEG C of indoor conditionss, injects the nutrient solution of aequum in culture vessel, and 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 running 4 days continuously.
C to take a morsel liquid from culture vessel liquid outlet, examines under a microscope counting, judges whether to satisfy the demands.
In steps A, described algae initial inoculation concentration at least need reach 10
4individual/mL, pollution-free for the algae inoculated, reach stable logarithmic phase.
In step B, described light source deceleration loading device ran than 16:8 according to the light dark time.
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, and structure is simple, manufacture and easy to maintenance.2, culture vessel that the present invention adopts is once-forming and volume is little, avoids the pollution of other material and is easy to change, facilitating succeeding transfer culture; Algae single culture vessel 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 voluntarily according to the different demands of cultivation algae kind to light intensity, and multiple algae can be cultivated simultaneously.3, after off-test, solution directly after sterilizing, can be poured out by liquid outlet, the workload reduced a staff.
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 only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
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, present embodiment discloses algae secondary culture apparatus, comprise bottom support plate, form framework at the upper fixing reflex reflector 2 of bottom support plate, back up pad preferentially selects stainless steel; The tool hole removable cover 1 that reflex reflector 2 carries, tool hole removable cover just covers reflex reflector framework; Tool hole removable cover arranges gas-loaded device 4; Light source deceleration loading device 3 and algae culture vessel 5 is provided with in framework; Mobile pulley 6 is installed bottom reflex reflector.The synthetic glass that described tool hole removable cover 1 is fixed for length and width, digs diameter 3cm circular hole in long edge and evenly arranges, each 3 of both sides.
As shown in Figure 2, light source deceleration loading device 3 comprises, be welded on the steelframe 20 in back up pad short side 16 mid-way, steelframe interposition installs several stainless steel columns 18, extend the buttress shaft 19 of band fixed ring 8 from wherein two stainless steel columns, other stainless steel column can according to cultivate algae kind need load varying strength light source 7, light source 7 is controlled by timing device, (namely open 16h daytime, close 8 hours night) is run than 16:8 according to the light dark time.Fixed ring 8 is apart from light source >=30cm, and the upper leaving certain gaps uncovered by the economic plan of circle, be convenient to picking and placeing of algae culture vessel 5; Reflex reflector is only fixed whole of back up pad long side, and surface is " Z " shape fold, guarantees uniform light.
As shown in Figure 3, gas-loaded device comprises air volume regulating valve 11 and gas take 10.Air volume regulating valve 11 regulates the gas velocity be charged in culture vessel by knob; Gas take 10 liang of side joint silicone tubes connect air pump and culture vessel, therebetween 0.22um filter membrane.
As shown in Figure 4, culture vessel 4 is that bottom and sidewall seal, the container of 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.Volume markings is carved with outside culture vessel 5 wall.Inlet pipe 12 passes through to bottom culture vessel 5 in the middle of tampon 14.
The present embodiment is simple and easy to do, workable, and material therefor is easy to obtain, and structure is simple, manufacture and easy to maintenance.Culture vessel that the present embodiment adopts is once-forming and volume is little, avoids the pollution of other material and is easy to change, 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 pollute, and ensure that the science of all kinds of research.Middle amount algae culture will be made to be convenient to carry out for the application of the present embodiment and purity is ensured.
Embodiment 2:
Utilize above-mentioned device to carry out a cultural method for conventional algae, the method comprises the steps:
A, under 25 ± 2 DEG C of indoor conditionss, injects the nutrient solution of 2L in culture vessel, and 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 inoculated, reach stable logarithmic phase.
B culture vessel fixes in algatron, open light source deceleration loading device, gas-loaded device, continuous operation observed algae density after 4 days, light source deceleration loading device described in this process ran than 16:8 according to the light dark time, described gas-loaded device, adds load volume for forming a string bubble.
C to take a morsel liquid from culture vessel liquid outlet, examines under a microscope counting, judges whether to meet work requirements.
Embodiment 3:5 kind commonly uses 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 conveniently can obtain the pure culture liquid of many algaes kind, meet the demand of research 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: temperature 23 ± 2 DEG C, 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 corresponding mother liquid concentration, and according to indicating that corresponding mother liquor consumption is transferred in 1000mL volumetric flask by order successively, constant volume, through autoclaving (121 DEG C, 15min), seals and posts label, 4 DEG C of Refrigerator stores, validity period 2 months.This substratum can use with after the distilled water diluting 10 times of autoclaving (121 DEG C, 15min).
* 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 to be dissolved in the 1mol/L HCl that more than 10mL step prepared, the 0.1mol/L EDTA-Na2 then 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 porous plug seals, boiling water heating 3h in a water bath, cooling, precipitation 24h, this process carries out 3 times continuously, then filter, get supernatant liquor, save backup in 4 DEG C of refrigerators after sterilizing in high-pressure sterilizing pot.
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, every 4d switching once.Culture transferring 2 ~ 3 times, treats that frustule is in logarithmic phase and reaches synchronous growth, namely can be used for enlarged culturing.
4 secondary enlarged culturing methods
Get the algae kind 20mL that preculture is 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.Get algae kind in culture vessel every 24h and measure frustule number.
5 cultivation results
Measure the algae density value of each treatment group with blood counting chamber, each nutrient solution at least counts twice, obtains mean value.If twice count results difference is comparatively large, then 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 5 class algae kinds used all energy normal growths, and cell count rate of growth is good, can practical requirement.
Claims (1)
1. utilize algae secondary culture apparatus to carry out a cultural method for conventional algae, it is characterized in that,
Described algae secondary culture apparatus comprises, bottom support plate, framework is formed at the upper fixing reflex reflector (2) of bottom support plate, in upper tool hole removable cover (1) of carrying of reflex reflector (2), tool hole removable cover (1) is arranged gas-loaded device (4), in framework, be 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), 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) is opened wide;
Described light source deceleration loading device (3) comprises the vertical steelframe (20) being arranged on the short side of back up pad (16) mid-way, the stainless steel column (18) being arranged on steelframe mid-way, and the buttress shaft (19) of the band fixed ring (8) to extend from stainless steel column, stainless steel column is arranged light source (7); Described fixed ring is for placing described algae culture vessel (5);
Described gas-loaded device (4) comprises air pump and connects the inlet pipe (12) of 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);
The inner surface of reflector is set to fold face;
Described fixed ring (8) distance light source (7) more than distance 30cm;
Cultural method comprises the steps:
A, under 25 ± 2 DEG C of indoor conditionss, inject nutrient solution in algae culture vessel (5), nutrient solution is tap water or substratum, and 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 running 3 ~ 4 days continuously;
C, get about 5ml liquid from algae culture vessel (5) liquid outlet, examine under a microscope counting, judge whether to meet the requirement reaching logarithmic phase, require index be operation after 4 days algae liquid concentration reach 10
6individual/more than mL;
In step B, described light source deceleration loading device (3) ran than 16:8 according to the light dark time;
Described gas-loaded device (4) loads demand 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 |
| 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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| CN203333661U (en) * | 2013-06-14 | 2013-12-11 | 金华职业技术学院 | Microalgae spread cultivation device |
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| CN101265449A (en) * | 2007-03-14 | 2008-09-17 | 中国科学院大连化学物理研究所 | Fast high-density culture method for algae cell |
| CN201395597Y (en) * | 2009-05-06 | 2010-02-03 | 向威 | Device for fast cultivating microalgae |
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