CN110437973A - A kind of culture apparatus and cultural method of anaerobic bacteria - Google Patents
A kind of culture apparatus and cultural method of anaerobic bacteria Download PDFInfo
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The present invention relates to a kind of culture apparatus of anaerobic bacteria and cultural methods, the device includes heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature controller, temperature probe, heated probe, measurement of oxygen content device, measurement of oxygen content probe, liquid storage bottle is placed in heat insulation trough, the bottleneck of liquid storage bottle is equipped with bottle stopper, and exhaust pipe, liquid injection pipe sequentially pass through bottle stopper and stretch in liquid storage bottle；Exhaust pipe stretches to the position in liquid storage bottle more than the liquid level of liquid；Liquid injection pipe stretches to liquid storage bottle bottom；Heat insulation trough side is equipped with temperature controller, and temperature controller is connected with temperature probe and heated probe, and temperature probe, heated probe probe into heat insulation trough through heat insulation trough side wall；Measurement of oxygen content probe is placed in bottle stopper bottom, and measurement of oxygen content probe is connect by passing through the cable of bottle stopper with measurement of oxygen content device.Overcome in the prior art that structure is complicated for culture device for anaerobic bacteria, can not overall process provide needed for gaseous environment defect.
Description
Technical field
The application belongs to the technical field of microculture, and in particular to a kind of culture device for anaerobic bacteria and culture side
Method.
Background technique
The culture of microorganism needs four big indispensable primary conditions: culture medium (that is, nutriment), temperature, gaseous environment
And pH value.Anaerobic bacteria (anaerobic bacteria) is a kind of thinner than what is grown in aerobic environment under anaerobic
Bacterium, and cannot be grown in the solid culture primary surface under air (oxygen containing 18%) and/or 10% gas concentration lwevel thin
Bacterium.It is increasing to the pure culture demand of anaerobic bacteria with the continuous development of biotechnology.
Currently, the culture device of anaerobic bacteria includes anaerobism bag, anaerobic jar and anaerobism work station.However, anaerobism bag, anaerobic jar
Corresponding gaseous environment can only be provided to the sample of cultivation stage, the gaseous environment of operational phase can not be but provided, along with behaviour
Make the reasons such as inconvenience, consumables cost height, and gradually tends to superseded.And the anaerobism work station that can cultivate overall process formula can be both operated
Have the defects that buying with use it is at high cost, take up space it is excessive.
Therefore, be badly in need of designing that a kind of structure is simple, it is small to take up space, and be capable of provided in overall process needed for gaseous environment
The culture apparatus of small-sized anaerobic bacteria, to overcome the above problem in the prior art.
Summary of the invention
The purpose of the present invention is to provide a kind of small-sized culture device for anaerobic bacteria, overcome anaerobic bacteria culture in the prior art
Apparatus structure is complicated, can not overall process provide needed for gaseous environment defect.To reach the purpose, the technical side that the present invention uses
Case is:
A kind of culture apparatus of anaerobic bacteria, including heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature controller,
Temperature probe, heated probe, measurement of oxygen content device, measurement of oxygen content probe, liquid storage bottle are placed in heat insulation trough, liquid storage bottle
Bottleneck is equipped with bottle stopper, and exhaust pipe, liquid injection pipe sequentially pass through bottle stopper and stretch in liquid storage bottle；Exhaust pipe stretches to liquid in liquid storage bottle
Ullage position；Liquid injection pipe stretches to liquid storage bottle lower part；Heat insulation trough side is equipped with temperature controller, and temperature controller is connected with survey
Temperature probe and heated probe, temperature probe, heated probe probe into heat insulation trough through heat insulation trough side wall；Measurement of oxygen content
Probe is placed in bottle stopper bottom, and the measurement of oxygen content probe is connect by passing through the cable of bottle stopper with measurement of oxygen content device.
Preferably, the culture apparatus of the anaerobic bacteria further includes air filter；The air filter and institute
State exhaust pipe connection.
Preferably, the temperature probe is resistance temperature measurement meter, thermoelectric couple thermo detector or pressure type thermo detector；Heating is visited
Head is electromagnetic heating rod or resistance heating rod；The temperature controller is digital temperature controller.
Preferably, being equipped with pipeline valve on the liquid injection pipe, exhaust pipe；The exhaust pipe connection aspiration pump or vacuum
Pump.
Another object of the present invention is to provide a kind of cultural method of anaerobic bacteria, can shorten the domestication training of anaerobic bacteria
The time is supported, high biochemical treatment efficiency is provided, is included the following steps:
(1), it sterilizes to heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature probe, heated probe
Processing；
(2), preparation culture sample, the culture bacterium of quasi- Anaerobic culturel be inoculated with spare；
(3), the bottle stopper for being connected with exhaust pipe, liquid injection pipe is covered after culture solution being added into liquid storage bottle；Guarantee the exhaust pipe
Stretch to the position in the liquid storage bottle more than the liquid level of liquid；The liquid injection pipe stretches to the liquid storage bottle bottom of bottle；
(4), above-mentioned culture sample is injected into culture solution using syringe；Nutrient solution is injected into culture solution；It extracts mixed
There is the culture solution of culture sample to detect its pH value；
(5), heated probe is controlled by temperature controller to be heated according to preset temperature, measured according to temperature probe
Real time temperature is adjusted；
(6), it is discharged using the air in aspiration pump or vacuum pump liquid storage bottle via exhaust pipe；
(7), temperature controller display carries out constant temperature timing culture after reaching preset temperature.
Preferably, the ingredient of the nutrient solution is by weight are as follows: 30 parts of sodium chloride, 20 parts of ammonium chloride, 10 parts of magnesium sulfate,
5 parts of potassium dihydrogen phosphate, 20 parts of morpholino b acid, 10 parts of yeast extract.
Preferably, the ingredient of the fermentation culture is by weight are as follows: 10 parts of glucose, 20 parts of sodium chloride, penicillin
10 parts, 5 parts of aspergillus niger, 5 parts of dipotassium hydrogen phosphate, 10 parts of zinc sulfate, 10 parts of ammonium sulfate, 5 parts of citric acid, 10 parts of yeast extract, morphine
5 parts of ethanesulfonic acid.
Preferably, condition of culture are as follows: preset temperature is 30-50 DEG C, pH value 3-7,50-70 hour.
Preferably, the air filter includes three-stage filtration, the first order is filtered into the absorbent cotton bucket of sterilizing, the
Two, three-stage filtration is the degreasing swab stick or ball of sterilizing.
The beneficial effects of the present invention are: being arranged using liquid storage bottle, exhaust pipe, syringe etc., so that anaerobic bacteria obtains abundance
Nutrition；It is arranged using heat insulation trough, temperature controller, temperature probe, heated probe etc., required cultivation temperature condition is carried out stringent
Control；Setting measurement of oxygen content device, measurement of oxygen content probe are measured in real time oxygen content；Aspiration pump or vacuum are set
Pump and filter device, are discharged, so that the exhaust gas in culture apparatus emits under condition of negative pressure, together after gas filtration
Shi Buhui pollutes culture.It has cheap, simple and practical, the controllable advantage of culture environment.Using the present invention
Culture device for anaerobic bacteria, convenient for laboratory student and small scale producers and experimenter's practice or use, solve small-scale raw
The problem of production person injects capital into.In method and step, using syringe for be inoculated with strain, drawing liquid detection, supplement the nutrients and etc.,
It is easy to operate, improve production efficiency；Culture solution and activator component are optimized, culture solution of the invention guarantees biochemical bacteria
Normal growth, promote the domestication of anaerobic species, shorten the domestication time of anaerobic species, improve biochemical treatment efficiency；The present invention
Nutrient solution improve anaerobic species activity, promote anaerobic species breeding, prevent the generation of water oxygen.Using anaerobism of the invention
Bacterium culture apparatus convenient for laboratory student and small scale producers and experimenter practice or uses, solves small scale producers and throw
The problem of entering fund.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of culture device for anaerobic bacteria of the invention.
Fig. 2 is digital temperature controller circuit theory schematic diagram of the invention.
Wherein: 1, heat insulation trough；2, liquid storage bottle；3, bottle stopper；4, exhaust pipe；5, liquid injection pipe；6, pipeline valve；7, it injects
Device；8, temperature controller；9, temperature probe；10, heated probe；11, measurement of oxygen content device；12, measurement of oxygen content is popped one's head in.
Specific embodiment
It is described in detail below in conjunction with specific embodiment of the attached drawing to technical scheme, to help this field skill
Art personnel are more fully understood by the technical solution of the application.
Embodiment one
A kind of culture apparatus of anaerobic bacteria, including heat insulation trough 1, liquid storage bottle 2, exhaust pipe 4, liquid injection pipe 5, syringe 7, control
Warm instrument 8, temperature probe 9, heated probe 10, measurement of oxygen content device 11, measurement of oxygen content probe 12.Liquid storage bottle 2 is placed in heat preservation
In sink 1, the bottleneck of liquid storage bottle 2 is equipped with bottle stopper 3, and exhaust pipe 4, liquid injection pipe 5 sequentially pass through bottle stopper 3 and stretch in liquid storage bottle 2；Row
Tracheae 4 stretches to the position in liquid storage bottle 2 above liquid levels；Liquid injection pipe 5 stretches to 2 bottom of liquid storage bottle；1 side of heat insulation trough
Equipped with temperature controller 8, temperature controller 8 is connected with temperature probe 9 and heated probe 10, and temperature probe 9, heated probe 10 are through heat preservation
The side wall of sink 1 probes into heat insulation trough 1；Measurement of oxygen content probe 12 is positioned at the lower end of bottle stopper 3, measurement of oxygen content probe
12 are connect by passing through the cable of bottle stopper 3 with measurement of oxygen content device 11.
The culture apparatus of the anaerobic bacteria further includes air filter (not shown)；The air filter with
The exhaust pipe 4 connects.
The temperature probe 9 is resistance temperature measurement meter；Heated probe 10 is electromagnetic heating rod；The temperature controller 8 is digital
Temperature controller, Fig. 2 shows its circuit theory schematic diagrams.
Preferably, being equipped with pipeline valve on the liquid injection pipe 5, exhaust pipe 4；The exhaust pipe 4 connects aspiration pump (figure
In be not shown).
Embodiment two
A kind of culture apparatus of anaerobic bacteria, including heat insulation trough 1, liquid storage bottle 2, exhaust pipe 4, liquid injection pipe 5, syringe 7, control
Warm instrument 8, temperature probe 9, heated probe 10, measurement of oxygen content device 11, measurement of oxygen content probe 12.Liquid storage bottle 2 is placed in heat preservation
In sink 1, the bottleneck of liquid storage bottle 2 is equipped with bottle stopper 3, and exhaust pipe 4, liquid injection pipe 5 sequentially pass through bottle stopper 3 and stretch in liquid storage bottle 2；Row
Tracheae 4 stretches to the position in liquid storage bottle 2 more than the liquid level of liquid；Liquid injection pipe 5 stretches to 2 bottom of liquid storage bottle；Heat insulation trough 1 one
Side is equipped with temperature controller 8, and temperature controller 8 is connected with temperature probe 9 and heated probe 10, and temperature probe 9, heated probe 10 are through guarantor
1 side wall of warm water tank probes into heat insulation trough 1；Measurement of oxygen content probe 12 is placed in 3 bottom of bottle stopper, the measurement of oxygen content probe 12
Cable by passing through bottle stopper 3 is connect with measurement of oxygen content device 11.
The culture apparatus of the anaerobic bacteria further includes air filter (not shown)；The air filter with
The exhaust pipe 4 connects.
The temperature probe 9 is thermoelectric couple thermo detector；Heated probe 10 is resistance heating rod.The temperature controller 8 is number
Word formula temperature controller, Fig. 2 shows its circuit theory schematic diagrams.
Preferably, being equipped with pipeline valve on the liquid injection pipe 5, exhaust pipe 4；The exhaust pipe 4 connects vacuum pump (figure
In be not shown).
Embodiment three
A kind of culture apparatus of anaerobic bacteria, including heat insulation trough 1, liquid storage bottle 2, exhaust pipe 4, liquid injection pipe 5, syringe 7, control
Warm instrument 8, temperature probe 9, heated probe 10, measurement of oxygen content device 11, measurement of oxygen content probe 12, liquid storage bottle 2 is placed in heat preservation
In sink 1, the bottleneck of liquid storage bottle 2 is equipped with bottle stopper 3, and exhaust pipe 4, liquid injection pipe 5 sequentially pass through bottle stopper 3 and stretch in liquid storage bottle 2；Row
Tracheae 4 stretches to the position in liquid storage bottle 2 more than the liquid level of liquid；Liquid injection pipe 5 stretches to 2 bottom of liquid storage bottle；Heat insulation trough 1 one
Side is equipped with temperature controller 8, and temperature controller 8 is connected with temperature probe 9 and heated probe 10, and temperature probe 9, heated probe 10 are through guarantor
1 side wall of warm water tank probes into heat insulation trough 1；Measurement of oxygen content probe 12 is placed in 3 bottom of bottle stopper, the measurement of oxygen content probe 12
Cable by passing through bottle stopper 3 is connect with measurement of oxygen content device 11.
The culture apparatus of the anaerobic bacteria further includes air filter (not shown)；The air filter with
The exhaust pipe 4 connects.
The temperature probe 9 is pressure type thermo detector；Heated probe 10 is resistance heating rod.The temperature controller 8 is number
Formula temperature controller, Fig. 2 shows its circuit theory schematic diagrams.
Preferably, being equipped with pipeline valve 6 on the liquid injection pipe 5, exhaust pipe 4；The exhaust pipe 4 connects vacuum pump
(not shown).
Using a kind of cultural method of anaerobic bacteria of above-mentioned culture device for anaerobic bacteria, the domestication culture of anaerobic bacteria can be shortened
Time provides high biochemical treatment efficiency, includes the following steps:
(1), it sterilizes to heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature probe, heated probe
Processing；
(2), preparation culture sample, the culture bacterium of quasi- Anaerobic culturel be inoculated with spare；
(3), the bottle stopper for being connected with exhaust pipe, liquid injection pipe is covered after culture solution being added into liquid storage bottle；Guarantee the exhaust pipe
Stretch to the position in the liquid storage bottle more than the liquid level of liquid；The liquid injection pipe stretches to the liquid storage bottle bottom of bottle；
(4), above-mentioned culture sample is injected into culture solution using syringe；Nutrient solution is injected into fermentation culture；It takes out
The culture solution for being mixed with culture sample is taken to detect its pH value；
(5), heated probe is controlled by temperature controller to be heated according to preset temperature, measured according to temperature probe
Real time temperature is adjusted；
(6), it is discharged using the air in aspiration pump or vacuum pump liquid storage bottle via exhaust pipe；
(7), temperature controller display carries out constant temperature timing culture after reaching preset temperature.
The ingredient of nutrient solution is by weight are as follows: 30 parts of sodium chloride, 20 parts of ammonium chloride, 10 parts of magnesium sulfate, potassium dihydrogen phosphate 5
Part, 20 parts of morpholino b acid, 10 parts of yeast extract.
The ingredient of fermentation culture is by weight are as follows: 10 parts of glucose, 20 parts of sodium chloride, 10 parts of penicillin, aspergillus niger 5
Part, 5 parts of dipotassium hydrogen phosphate, 10 parts of zinc sulfate, 10 parts of ammonium sulfate, 5 parts of citric acid, 10 parts of yeast extract, 5 parts of morphine ethanesulfonic acid.
Condition of culture are as follows: preset temperature is 30-50 DEG C, pH value 3-7,50-70 hour.
Air filter includes three-stage filtration, and the first order is filtered into the absorbent cotton bucket of sterilizing, second and third grade, which is filtered into, goes out
The degreasing swab stick or ball of bacterium.
What has been described above is only a preferred embodiment of the present invention, it is noted that for those skilled in the art,
Without depart from that overall concept of the invention, several changes and improvements can also be made, these also should be considered as of the invention
Protection scope.
Claims (9)
1. a kind of culture apparatus of anaerobic bacteria, including heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature controller, survey
Warm probe, heated probe, measurement of oxygen content device, measurement of oxygen content probe, which is characterized in that liquid storage bottle is placed in heat insulation trough
Interior, the bottleneck of liquid storage bottle is equipped with bottle stopper, and exhaust pipe, liquid injection pipe sequentially pass through bottle stopper and stretch in liquid storage bottle；Exhaust pipe stretches to
Position in liquid storage bottle above liquid levels；Liquid injection pipe stretches to liquid storage bottle liquid lower part；Heat insulation trough side is equipped with temperature controller,
Temperature controller is connected with temperature probe and heated probe, and temperature probe, heated probe probe into heat preservation water through heat insulation trough side wall
In slot；Measurement of oxygen content probe is placed in bottle stopper following side, the measurement of oxygen content probe by pass through the cable of bottle stopper with it is oxygen-containing
Amount detecting device connection.
2. the culture apparatus of anaerobic bacteria according to claim 1, which is characterized in that the culture apparatus of the anaerobic bacteria also wraps
Include air filter；The air filter is connect with the exhaust pipe.
3. the culture apparatus of anaerobic bacteria according to claim 1, which is characterized in that the temperature probe is resistance temperature measurement
Meter, thermoelectric couple thermo detector or pressure type thermo detector；Heated probe is electromagnetic heating rod or resistance heating rod；The temperature controller is
Digital temperature controller.
4. the culture apparatus of anaerobic bacteria according to claim 1, which is characterized in that be all provided on the liquid injection pipe, exhaust pipe
There is pipeline valve；The exhaust pipe connection aspiration pump or vacuum pump.
5. a kind of cultural method of anaerobic bacteria, which is characterized in that using the training of anaerobic bacteria of any of claims 1-4
Device is supported, is included the following steps:
(1), sterilization treatment is carried out to heat insulation trough, liquid storage bottle, exhaust pipe, liquid injection pipe, syringe, temperature probe, heated probe；
(2), preparation culture strain, the culture bacterium of quasi- Anaerobic culturel be inoculated with spare；
(3), the bottle stopper for being connected with exhaust pipe, liquid injection pipe is covered after fermentation culture being added into liquid storage bottle；Guarantee the exhaust pipe
Stretch to the position in the liquid storage bottle more than the liquid level of liquid；The liquid injection pipe stretches to the liquid storage bottle bottom of bottle；
(4), above-mentioned culture strain is injected into fermentation culture using syringe；Nutrient solution is injected into fermentation culture；It takes out
The culture solution containing anaerobic bacteria is taken to detect its pH value；
(5), heated probe is controlled by temperature controller to be heated according to preset temperature, measured according to temperature probe real-time
Temperature is adjusted；
(6), the exhaust gas in liquid storage bottle is discharged via exhaust pipe using aspiration pump or vacuum pump；
(7), temperature controller display carries out constant temperature timing culture after reaching preset temperature.
6. the cultural method of anaerobic bacteria according to claim 5, which is characterized in that the ingredient of the nutrient solution is by weight
Are as follows: 30 parts of sodium chloride, 20 parts of ammonium chloride, 10 parts of magnesium sulfate, 5 parts of potassium dihydrogen phosphate, 20 parts of morpholino b acid, 10 parts of yeast extract.
7. the cultural method of anaerobic bacteria according to claim 5, which is characterized in that the ingredient of the fermentation culture is by weight
Meter are as follows: 10 parts of glucose, 20 parts of sodium chloride, 10 parts of penicillin, 5 parts of aspergillus niger, 5 parts of dipotassium hydrogen phosphate, 10 parts of zinc sulfate, sulphur
10 parts of sour ammonium, 5 parts of citric acid, 10 parts of yeast extract, 5 parts of morphine ethanesulfonic acid.
8. the cultural method of anaerobic bacteria according to claim 5, which is characterized in that condition of culture are as follows: preset temperature 30-
50 DEG C, pH value 3-7,50-70 hour.
9. the cultural method of anaerobic bacteria according to claim 5, which is characterized in that air filter includes three-level mistake
Filter, the first order are filtered into the absorbent cotton bucket of sterilizing, second and third grade is filtered into the degreasing swab stick or ball of sterilizing.
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Cited By (4)
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| CN111562331A (en) * | 2020-06-10 | 2020-08-21 | 四川农业大学 | Improved nitrogenase activity assay device and assay method using acetylene reduction method |
| CN111925920A (en) * | 2020-08-14 | 2020-11-13 | 常州惠农生物科技有限公司 | Culture bottle for anaerobic bacteria fermentation |
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| CN111562331A (en) * | 2020-06-10 | 2020-08-21 | 四川农业大学 | Improved nitrogenase activity assay device and assay method using acetylene reduction method |
| CN111925920A (en) * | 2020-08-14 | 2020-11-13 | 常州惠农生物科技有限公司 | Culture bottle for anaerobic bacteria fermentation |
| CN117106577A (en) * | 2023-10-13 | 2023-11-24 | 北京万通益生物科技有限公司 | Lactic acid bacteria hot-drying method preparation environmental monitoring system based on thing networking |
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