CN105210844B - Utilize the simulation in-situ batch culture system of the tracer techniques of C 14 measure primary productivity of marine ecosystem - Google Patents
Utilize the simulation in-situ batch culture system of the tracer techniques of C 14 measure primary productivity of marine ecosystem Download PDFInfo
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- CN105210844B CN105210844B CN201510685748.4A CN201510685748A CN105210844B CN 105210844 B CN105210844 B CN 105210844B CN 201510685748 A CN201510685748 A CN 201510685748A CN 105210844 B CN105210844 B CN 105210844B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G33/00—Cultivation of seaweed or algae
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
- A01G31/02—Special apparatus therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/005—Incubators
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Abstract
The present invention relates to a kind of simulation in-situ batch culture system that primary productivity of marine ecosystem is determined using the tracer techniques of C 14, including culture unit, temperature and water flow simulation control unit, lighting simulation control unit；Culture unit includes several blake bottles；Temperature and water flow simulation control unit include water-bath cover, recirculated water bath, blake bottle runing rest, water-bath cover is cylindrical shape, center rotational shaft is set along axis in cylinder, center rotational shaft periphery sets blake bottle runing rest, blake bottle is fixed on blake bottle runing rest, water inlet and delivery port, water inlet and delivery port is set to be connected respectively with recirculated water bath on water-bath cover；Lighting simulation control unit includes optic panel, fluorescent tube, and fluorescent tube is arranged on optic panel, and fluorescent tube connection power supply, lighting simulation control unit is arranged on the bottom surface side of cylindrical water-bath cover.The system can accurately control water temperature, intensity of illumination, can simulate ocean dynamical environment, prevent C 14 from leaking.
Description
Technical field
The present invention relates to the culture systems of phytoplankton, and in particular to one kind is primary using C-14 tracer techniques measure ocean
The simulation in-situ batch culture system of productivity.
Background technology
Primary productivity of marine ecosystem refers to primary producer in ocean (mainly phytoplankton) by photosynthesis or chemistry
The ability or speed of synthetically produced organic matter, it is the existence basis of other heterotrophic organisms in marine ecosystems, and from basic
On affect Global Biogeochemical Cycle and climate change.Therefore, the accurate test of primary productivity of marine ecosystem has turned into each
The important pivot of carbon cycle, ocean carbon sequestration capacity and climate change between state's research active organism and inorganic carbon reservior.C-
14 tracer methods are primary productivity standard assay techniques the most classical and conventional so far, by Danish scientist
Steemann Nielsen propose in nineteen fifty-two, substitute the black of conventional prevalence rapidly because its accuracy is high, it is short to take, expense is low
White bottle oximetry, it is widely used by thalassographer and lakes and marhshes scholar, turn into International Standards Method at present.The technology is in actual behaviour
Two methods can be divided into work：" original position " in-situ batch culture method and " simulation " in-situ batch culture method, the former requires adding on blake bottle
Predetermined depth under water is placed on after water sample and C-14, and requires that investigating secure waits 4-6 hours (or more than 24 hours)
Until culture terminates, so difficulty is high, time-consuming and laborious；And the latter is then by engineering technology come simulated field conditions, so as to real
Now simulate the incubation of original position.
Although there is the technical method of " simulation " in-situ batch culture in the past, these technologies are present following aspects not
Foot part：
(1) temperature being difficult in accurate control incubation.All the time in change, this is not only showed ocean water body temperature
In the seasonal variations of large scale, the mechanical periodicity that day alternates with night in one day is also manifested by.Moreover, ocean water body has vertical solid
Dimension, different water depth temperature are also different.Simulation culture technique was just again difficult to control the temperature of water body after water sample is taken out in the past
Degree change, the temperature of the depth where temperature can not be more strict controlled in water sample originally.Research showed that temperature was to swimming in the past
Vegetative primary productivity and photosynthesis carbon assimilation index impacts are very big, and this certainly will cause the error of C-14 traces.
(2) intensity of illumination being difficult in accurate control incubation.Illumination be influence primary productivity of marine ecosystem it is crucial because
Element, because illumination is the motive power that phytoplankton produces organic carbon by photosynthesis.Although conventional partial simulation culture technique
The simulated solar illumination condition such as fluorescent lamp is employed, but is often difficult to the intensity of precision control illumination, it is difficult to be former deep with water sample
The intensity of illumination of degree is united；Even if part culture technique can adjust fluorescent lamp intensity of illumination, but due to not being fully sealed
Environment, cause ambient natural light shine on phytoplankton produce be difficult to expected influence, and then cause test result error and
It is uncertain.
(3) it is difficult to analog physical hydrodynamic environment.The ocean water body moment is in motion, and this physical motion is to swimming
Vegetative primary productivity has considerable influence.Simulation culture technique was often again difficult after water sample is taken out from deep-sea in the past
To simulate phytoplankton script possessed ocean dynamical environment.This will make it that surveyed primary productivity and legitimate reading are in distress
With expected deviation.
(4) it is difficult to radioactive pollution caused by leaking C-14 strictly to be controlled.Conventional culture technique is noted by C-14
Inject in blake bottle, and incubation, or even be all difficult to avoid that caused by manual operation during last filtering water sample
C-14 is leaked and radioactive pollution.Because C-14 is radio isotope, and its long half time, up to 5730 or so, this causes C-
14 pass to the mankind possibly through food chain, and then influence human health.On the other hand, due to being injected during culture in water sample
C-14 concentration is generally large, if any leakage, also easily pollutes research ship, it is difficult to remove；If large batch of persistently leakage, also has
Increasing for ocean background C-14 contents may be caused.
In a word, conventional art is difficult to the perfect real site environment of simulation, so as to cause measurement result to be deviated and lose
By mistake, and radioactive pollution is easily caused.Simulation to site environment is included to multiple ginsengs such as temperature, intensity of illumination, physical perturbation
Several perfect reproductions, but conventional art method is often only capable of simulating 1-2 parameter therein, it is difficult to all conditions are all strictly controlled
System；Especially under the research ship confined space and dynamic environment, the strict control to temperature, the accurate reproduction to physical perturbation and right
The strict simulation of luminous intensity all runs into extreme difficulty；In addition, how to reduce C-14 leakages and the radioactivity caused by manual operation
Pollution；This is all the difficult point that conventional multiple technologies are difficult to capture.
The content of the invention
The present invention is intended to provide a kind of " simulation " in-situ batch culture system, to solve, existing culture systems operation difficulty is big, consumes
Duration, operation require high, easily cause the problems such as radioactive pollution；Meanwhile the high-precision feature of C-14 tracer techniques is made full use of,
Perfection solves in scientific investigation shipboard measurement primary productivity of marine ecosystem and carried out the problem of related science experiment, is adjusted for Marine Sciences
Look into and study and technical support is provided.
A kind of simulation in-situ batch culture system that primary productivity of marine ecosystem is determined using C-14 tracer techniques, including culture are single
Member, temperature and water flow simulation control unit, lighting simulation control unit；Culture unit includes several blake bottles；Temperature and water
Flow field simulation control unit includes water-bath cover, recirculated water bath, blake bottle runing rest, and water-bath cover is cylindrical shape, in cylinder
Body sets center rotational shaft along axis, and center rotational shaft periphery sets blake bottle runing rest, and blake bottle is fixed on blake bottle
On runing rest, water inlet and delivery port, water inlet and delivery port is set to be connected respectively with recirculated water bath on water-bath cover；Illumination
Analog controlling unit includes optic panel, fluorescent tube, and fluorescent tube is arranged on optic panel, fluorescent tube connection power supply, illumination
Analog controlling unit is arranged on the bottom surface side of cylindrical water-bath cover.
The quantity of the blake bottle is 4-10, can be needed to be adjusted the quantity of blake bottle according to actual experiment, one
As at least three transparent material blake bottle, " black " blake bottle use of 1-3 appearance blacking or jealous glass material can be included in addition
In doing dark absorption correction, two kinds of blake bottles can be with cross-distribution.
It is symmetrical arranged between the blake bottle by symmetry axis of center rotational shaft, makes its trim, is avoided unstable when rotated.
The culture bottleneck is arranged on the outside of water-bath cover.
The blake bottle is set for angle, i.e., bottleneck direction has angle with body direction, and unconventional direction is consistent.
The bottleneck of blake bottle is made into oblique, and be fixed on blake bottle runing rest, it is convenient water sample to be added into bottle, conveniently with injection
Device is injected into C-14 solution, the cleaning being also convenient for after culture；Cylindrical water-bath cover can be opened to come more from cylinder bottom surface
Blake bottle is changed, the quantity of blake bottle is such as adjusted, changes different light transmittances, the blake bottle of volume；And in incubation, training
Support bottle and be in closed state.
The water-bath cover is made of transparent material, and such as glass, high-transmittance plastics, transparent material do not interfere with illumination
Injection.
The water inlet and delivery port are arranged on the side of cylindrical water-bath cover, both at a distance of more remote better, make from
The water of recirculated water bath input circulates completely in water-bath cover.
Light quantum meter is set between the water-bath cover and lighting simulation control unit, and light quantum meter is close to water-bath cover, and position
In using the cylindrical water-bath cover bottom surface center of circle as on the circumference of the center of circle, the center of circle to blake bottle middle part distance for the circle of radius.
Light quantum meter is connected with computer, for detecting luminous intensity at any time, to ensure that Optical power values are accurate.
Current controller and/or timer can also be set between the fluorescent tube and power supply.
The external side of the center rotational shaft, installs detachable crank.Under the crank can be dismantled in incubation
Come, and before culture plus during water sample and after incubation wash bottle when, can install and easily shake, to cause on blake bottle
Lower rotation, so as to which easily current are poured into or poured out.Described Ke Chaixieshi can the conventional company such as be threadedly coupled, snap connection
Connect mode.The external side refers to the side that culture bottleneck leans out water-bath cover.
The water-bath cover is fixed on support.
The culture systems are arranged in addition to recirculated water bath in lighttight light shield.To avoid natural light outside line
Interference, using totally enclosed lighttight outside light shield, and the drawing curtain that light shield is used above light tight and closed, this
Sample can both play the effect of closing illumination in incubation, can also draw the curtain apart after incubation to be sampled.Outside hides
Light shield uses rigid, so when rocking or moving on scientific surveying ship, can protect the transparent water steam-inflated plastic shroud of the inside.Circulation
The reason for water-bath is placed on outside light shield is：First, saving the space of light shield, two are placed on outside easily radiating, third, side
Convenient to operate.
The light shield uses rigid, and body is tetragonal body, therefore conveniently cumulative can stack；Meanwhile if pin
The water sample of different water depth is cultivated, can be stacked together with this multiple kind equipment, and then is saved shared on research ship
Space.
Compared to prior art, beneficial effects of the present invention are embodied in：
(1) used for the shortcomings that being difficult to accurately control temperature in incubation, temperature simulation control unit of the invention
Circular non-opaque water-bath cover provides a totally-enclosed culture environment for blake bottle, and by circulator bath device drives from blake bottle
The constant temperature current flowed continuously through outside, only ± 0.1 DEG C of temperature control error；Moreover, the temperature control scope of water-bath is covered between 0 DEG C -100 DEG C
The viable temperature range of nearly all marine phytoplankton institute is covered.Therefore, the present invention can not only accurately simulated water sample in sea
The water temperature of the former depth in ocean, and suitable for any maritime waters in the whole world.
(2) for the shortcomings that being difficult to accurately control intensity of illumination in incubation, lighting simulation control unit of the invention
Using current intensity controller and timer control fluorescent tube and optic panel, so as to strictly control intensity of illumination and illumination mould
Formula, and pass through the further close inspection luminous intensity of light quantum meter, it is ensured that the luminous intensity of luminous intensity and water sample in former ocean water body
It is completely the same；Moreover, blake bottle is high transparency material with the cylindrical water-bath cover outside it, light transmission is not interfered with,
Ocean intensity of illumination can perfectly be simulated.In addition, it is a little to take outside light shield entirely to cultivate that the present invention is the most key
Equipment is all closed (except circulator bath equipment), so effectively reduces in surrounding environment natural lighting to blake bottle
Influence, so that test result is more accurate.
(3) for the shortcomings that being difficult to analog physical hydrodynamic environment, the current that the present invention is driven using circulator bath push away
Dynamic blake bottle persistently rotates around center rotational shaft, so as to simulate ocean water body physical motion；And blake bottle is in itself in flow action
Under not only passively receive current and bring thermostatic effect, and serve the effect for promoting runner, be to kill two birds with one stone.Water in bottle
Sample is because blake bottle rotates upwardly and downwardly, and in the presence of natural gravity, can mix up and down or side-to-side movement.In addition, pass through regulation
The flow velocity of constant temperature current controls the speed that blake bottle rotates, so as at utmost simulation ocean dynamical environment.
(4) it is directed to C-14 in conventional art and is easily compromised the shortcomings that causing radioactive pollution, the present invention passes through following several
Measure is improved：A. blake bottle bottleneck is changed to italic mouth and be fixed on runing rest, and added outside center rotational shaft
Add dismountable armstrong's patent crank, so can be by blake bottle rocking-turn to extreme lower position, then bottleneck upward, passes through injection
C-14 solution is injected into bottle by device, reduces potential C-14 escape of radioactivity.B., can be in exposed bottle after culture terminates
Conduit is put at mouthful, and blake bottle is shaken to inversion, liquid directly flows out to filter plant through conduit in bottle under gravity
Later stage filtering is carried out to facilitate, can so avoid the nutrient solution containing C-14 that other utensils are produced with radioactivity attachment pollution.C. exist
In whole incubation, culture bottleneck covering bottle cap, generally in full closeding state, so as to effectively prevent radioactive substance to let out
The generation of leakage.D. culture terminate after washing process in, it is only necessary to add cleaning solution, lid bottle cap, shake handle rotate washing,
With the conduit export operating procedure such as cleaning solution, without with hand or extremity radioactive substance, protecting health；Outflow
Cleaning solution directly imports waste liquid barrel through conduit, so as to reduce splashing in operation in the past or water clock, avoids to scientific investigation
The pollution of ship or surrounding environment.
Brief description of the drawings
Fig. 1 present system front views；
Fig. 2 present system side cutaway views；
Fig. 3 lighting simulation control unit structural representations.
In figure：1- water-bath covers；2- optic panels；3- light shields；4- center rotational shafts；5- blake bottles；6- blake bottle runing rests；
7- water inlets；8- oral siphons；9- recirculated water baths；10- outlet pipes；11- cultivates bottleneck；12- supports；13- delivery ports；14- shakes
；15- fluorescent tubes；16- current controllers；17- timers；18- power supplys；19- light quantum meters.
Embodiment
The present invention is described in further details below in conjunction with the accompanying drawings.
The simulation in-situ batch culture system that primary productivity of marine ecosystem is determined using C-14 tracer techniques as shown in Figure 1-2, bag
Include culture unit, temperature and water flow simulation control unit, lighting simulation control unit.
Culture unit includes six blake bottles 5, wherein have the blake bottle of three transparent materials, the training of three transparent materials
Support bottle.
Temperature and water flow simulation control unit include water-bath cover 1, recirculated water bath 9, blake bottle runing rest 6, water-bath cover 1
For cylindrical shape, center rotational shaft 4 is set along axis in cylinder, the periphery of center rotational shaft 4 sets blake bottle to rotate branch
Frame 6, blake bottle 5 are fixed on blake bottle runing rest 6, are symmetrical arranged between blake bottle 5 with center rotational shaft 4 for symmetry axis, training
The outside that bottleneck 11 is arranged on water-bath cover 1 is supported, sets water inlet 7 and delivery port 13, water inlet 7 to be arranged on water-bath on water-bath cover 1
Cover 1 top and the rotational trajectory of face blake bottle 5, delivery port 13 are arranged on the bottom of water-bath cover 1, water inlet 7 and delivery port
13 are connected by oral siphon 8 and outlet pipe 10 with recirculated water bath 9 respectively.
As shown in figure 3, lighting simulation control unit includes optic panel 2, fluorescent tube 15, fluorescent tube 15 is arranged on smooth surface
On plate 2, fluorescent tube 15, current controller 16, timer 17 and power supply 18 are sequentially connected, and lighting simulation control unit is arranged on
The bottom surface side of cylindrical water-bath cover 1.
Blake bottle 5 is set for angle, i.e., bottleneck direction has angle with body direction, and unconventional direction is consistent, sees
Shown in Fig. 2.The bottleneck of blake bottle 5 is made into oblique, and is fixed on blake bottle runing rest 6, it is convenient added into bottle water sample,
It is convenient to be injected into C-14 solution with syringe, be also convenient for the cleaning after cultivating；Cylindrical water-bath cover 1 can be from cylinder bottom
Face is opened to change blake bottle 5, is such as adjusted the quantity of blake bottle 5, is changed different light transmittances, the blake bottle 5 of volume；And
In incubation, blake bottle 5 is in closed state.
Water-bath cover 1 is made of high-transmittance plastics, does not interfere with the injection of illumination, and light non-friable, convenient to carry.
Light quantum meter 19 is set between water-bath cover 1 and lighting simulation control unit, light quantum meter 19 is close to water-bath cover 1, and
Positioned at using the bottom surface center of circle of cylindrical water-bath cover 1 as the circumference of the center of circle, the center of circle to the middle part distance of blake bottle 5 for the circle of radius
On.Light quantum meter 19 is connected with computer, for detecting luminous intensity at any time, to ensure that Optical power values are accurate.
The external side of center rotational shaft 4, namely culture bottleneck 11 lean out the side of water-bath cover 1, install detachable crank
14.The crank 14 can disassemble in incubation, and before culture plus during water sample and after incubation wash bottle when, can be with
Install and easily shake, to cause blake bottle 5 to rotate upwardly and downwardly, so as to which easily current are poured into or poured out.
Water-bath cover 1 is fixed on support 12.Whole culture systems are arranged at lighttight screening in addition to recirculated water bath 9
In light shield 3.To avoid the interference of natural light outside line, using totally enclosed lighttight outside light shield 3, and light shield 3
Used above light tight and closed drawing curtain, the effect of closing illumination so can be both played in incubation, can also trained
Drawn the curtain apart after supporting to be sampled.Outside light shield 3 uses rigid, so rocks or moves on scientific surveying ship
When, the transparent water steam-inflated plastic shroud 1 of the inside can be protected, the body of light shield 3 is tetragonal body, therefore conveniently cumulative can be stacked；Meanwhile
It if cultivated for the water sample of different water depth, can be stacked together, and then be saved on research ship with this multiple kind equipment
Shared space.Recirculated water bath 9 is placed on the reason for light shield 3 is outer and is：First, the space of light shield 3 is saved, second, putting
Easily radiating outside, third, conveniently operating.
Claims (7)
- A kind of 1. simulation in-situ batch culture system that primary productivity of marine ecosystem is determined using C-14 tracer techniques, it is characterised in that bag Include culture unit, temperature and water flow simulation control unit, lighting simulation control unit；Culture unit includes several blake bottles (5)；Temperature and water flow simulation control unit include water-bath cover (1), recirculated water bath (9), blake bottle runing rest (6), water-bath It is cylindrical shape to cover (1), center rotational shaft (4) is set along axis in cylinder, center rotational shaft (4) periphery sets culture Bottle runing rest (6), blake bottle (5) are fixed on blake bottle runing rest (6), and water inlet (7) is set on water-bath cover (1) and is gone out The mouth of a river (13), water inlet (7) and delivery port (13) are connected with recirculated water bath (9) respectively；Lighting simulation control unit includes light Panel (2), fluorescent tube (15), fluorescent tube (15) are arranged on optic panel (2), fluorescent tube (15) connection power supply (18), Lighting simulation control unit is arranged on the bottom surface side of cylindrical water-bath cover (1)；The blake bottle (5) sets for angle Put, culture bottleneck (11) is arranged on the outside of water-bath cover (1)；The external side of the center rotational shaft (4), installation is detachable to shake (14), the external side refers to the side that culture bottleneck (11) leans out water-bath cover (1)；The culture systems remove circulator bath It is arranged at outside pot (9) in lighttight light shield (3).
- 2. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, it is characterised in that the quantity of the blake bottle (5) is 4-10, with center rotational shaft (4) for symmetry axis between blake bottle (5) It is symmetrical arranged.
- 3. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, the water-bath cover (1) are made of transparent material.
- 4. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, sets light quantum meter (19) between the water-bath cover (1) and lighting simulation control unit, light quantum meter (19) is close to water-bath cover (1), and positioned at by the center of circle of cylindrical water-bath cover (1) the bottom surface center of circle, the center of circle to blake bottle middle part distance be radius On round circumference.
- 5. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, also sets up current controller (16) and/or timer (17) between the fluorescent tube (15) and power supply (18).
- 6. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, the water-bath cover (1) are fixed on support (12).
- 7. the simulation in-situ batch culture system according to claim 1 that primary productivity of marine ecosystem is determined using C-14 tracer techniques System, the light shield (3) use rigid, and body is tetragonal body.
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| KR100928088B1 (en) * | 2008-11-27 | 2009-11-23 | 주식회사 두산에코비즈넷 | A culture midium for microorganism |
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