Absolute anaerobic gas-producing or gas-consuming microorganism culture system

Abstract

The utility model discloses an absolutely anaerobic gas-producing or gas-consuming microorganism culture system which comprises a culture reactor connected with a liquid sampling pipeline, a gas outlet pipeline and a gas inlet pipeline, the liquid sampling pipeline and the gas inlet pipeline are connected with a two-way valve, the gas outlet pipeline is connected with a first pipeline and a second pipeline, and the first pipeline is connected with a third pipeline and a fourth pipeline. The fourth pipeline is connected with a pressure gauge, and the second pipeline is connected with a clamping sleeve right-angle valve; the air inlet pipeline and the seventh pipeline are both connected with an air storage tank, and the air storage tank is connected with a vacuum sampling pipeline; a sixth pipeline connected with an air inlet of the air pump is connected with the fourth pipeline and the fifth pipeline through a three-way valve, and a sixth pipeline connected with an air outlet of the air pump is connected with the seventh pipeline and the fifth pipeline through a three-way valve. According to the device disclosed by the utility model, gas phase and liquid phase sample collection and gas circulation in a microbial metabolism process are taken into consideration, and enrichment culture of microorganisms and anaerobic collection of microbial products under an absolutely anaerobic or anaerobic condition are facilitated.

Description

TECHNICAL FIELD

[0001] The utility model relates to anaerobic microbial culture system, concretely is a kind of absolute anaerobic gas production or gas consumption microbial culture system. BACKGROUND

[0002] Most of the traditional laboratory anaerobic culture system relies on the pre-treatment of culture medium to remove the dissolved oxygen in liquid medium or water-based medium, for example, the German Microorganism and Cell Culture Collection (DSMZ), one of the international authoritative institutions, often recommends using nitrogen and carbon dioxide mixed gas (v / v 8:2) to purge the sterilized medium for 30-45 minutes to remove the dissolved oxygen.

[0003] The existing anaerobic culture system often relies on small devices such as Hungate bottles or glass Schlenk flasks with butyl rubber caps and aluminum caps. The commonly used Hungate bottles and Schlenk flasks are colorless and transparent containers, which have the problem of culturing photosynthetic microorganisms. Once photosynthetic microorganisms produce oxygen, it will seriously affect the internal anaerobic environment. In addition, Hungate bottles and Schlenk flasks are made of glass and have small volume, which is not friendly to the culture of a large number of microorganisms in the field of environment and other related fields. In addition, the existing anaerobic culture system cannot simultaneously meet the needs of real-time gas replacement, real-time oxygen-free sampling of liquid samples, and anaerobic pressure culture simulation of river, lake and ocean environments. The invention patent with publication number CN112852614A "Anaerobic microbial culture medium oxygen removal device and oxygen removal method with adjustable gas composition" uses sealed Schlenk flasks and Hungate tubes as anaerobic containers. By controlling the gas flow, starting and stopping of the vacuum pump, and opening and closing of the electromagnetic valve through the controller, an oxygen-free microbial culture environment is created. However, due to the use of Schlenk flasks, real-time sampling and pressure culture cannot be achieved. The invention patent with publication number CN114958584A "Portable anaerobic culture device and its application" includes a box, a gas mixing and purification block, a gas filling and pumping block, and a gas injection block. The injection block continuously injects oxygen-free gas into the environment medium for oxygen removal pretreatment in an open system. The gas filling and pumping block replaces the gas in the culture bottle in a closed system. The system operation is not much different from conventional anaerobic gas replacement. It also involves the use of hypodermic needles and the frequent insertion and removal of needles through butyl rubber stoppers. The use of needles and butyl rubber stoppers results in poor sealing, and oxygen is inevitably introduced during the operation process, which cannot provide an absolute anaerobic environment. Moreover, the operation is complicated and requires high experimental skills of the operator. The invention patent with publication number CN108251299A "Anaerobic culture device and its application" uses nitrogen gas to purge and set up a gas collection bag connected to the culture container through a silica gel tube to create an anaerobic environment for in vitro intestinal flora culture. The sealing performance of the system is limited by the sealing performance of the gas collection bag connection. Since the gas collection bag is introduced, the system cannot withstand positive pressure. The invention patent with publication number CN209431090U "Anaerobic system based on vacuum pumping and gas filling" is mainly used for gas replacement in anaerobic culture systems and cannot collect samples under airtight conditions in the gas and liquid phases. The invention patent with publication number CN201176433Y "Anaerobic culture liquid preparation device" cannot ensure uniform gas distribution and does not consider the introduction of trace oxygen by the gas diffusion of the hose, resulting in poor anaerobic environment and complicated operation process.

[0004] In summary, it is not difficult to see that the existing anaerobic culture system mainly has the following problems: 1) little consideration is given to the collection of gas and liquid samples in the experiment / test / system operation; 2) the gas circulation in the system operation is not considered; 3) the existing anaerobic culture system often uses silica gel tubes, which are extremely easy to penetrate oxygen from the air due to diffusion, the air tightness of the system is poor during long-term operation, and the widely used subcutaneous injection needle, rotor flowmeter, air bag and other components restrict the anaerobic microbial culture under lower or higher than the atmospheric pressure. However, the gas pressure of the system is required in the research of river and lake mud microbial metabolism, marine environment sediment and deep sea biological activity, conventional gas production and microbial metabolism of gas substances. Therefore, it is necessary to develop a pressure-resistant, easy-to-sample gas and liquid sample, high-airtightness microbial culture and microbial product collection system. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide an absolute anaerobic gas-producing or gas-consuming microbial culture system, which takes into account the collection of gas and liquid samples and gas circulation, and is more conducive to the enrichment culture of microorganisms under absolute anaerobic or oxygen-free conditions.

[0006] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0007] An absolute anaerobic gas-producing or gas-consuming microbial culture system, comprising a culture reactor, a gas storage tank and a gas pump, the top of the culture reactor is respectively connected with a liquid sampling pipeline, a gas outlet pipeline and a gas inlet pipeline, and the liquid sampling pipeline and the gas inlet pipeline are both connected with a two-way valve;

[0008] The gas outlet pipeline is connected with a first pipeline and a second pipeline through a three-way valve, the first pipeline is connected with a third pipeline and a fourth pipeline through a three-way valve, the fourth pipeline is connected with a pressure gauge buffer pipeline through a three-way, and the pressure gauge buffer pipeline is connected with a pressure gauge, and the second pipeline is connected with a clamping sleeve right-angle valve;

[0009] The gas inlet pipeline is connected with the gas outlet of the gas storage tank, the gas inlet of the gas storage tank is connected with a seventh pipeline, the gas storage tank is connected with a vacuum sampling pipeline, and the vacuum sampling pipeline comprises a vertical pipeline section, a first horizontal pipeline section and a second horizontal pipeline section connected through a three-way valve, the vertical pipeline section is connected with a vacuum valve, and the second horizontal pipeline section is connected with a two-way valve;

[0010] The gas inlet and the gas outlet of the gas pump are respectively connected with a sixth pipeline, the sixth pipeline connected with the gas inlet of the gas pump is connected with the fourth pipeline and the fifth pipeline through a three-way valve, and the sixth pipeline connected with the gas outlet of the gas pump is connected with the seventh pipeline and the fifth pipeline through a three-way valve.

[0011] Further, the culture reactor, the gas storage tank, the liquid sampling pipeline, the gas inlet pipeline, the gas outlet pipeline, the first pipeline, the second pipeline, the third pipeline, the fourth pipeline, the fifth pipeline, the sixth pipeline, the seventh pipeline, the pressure gauge buffer pipeline and the vacuum sampling pipeline are made of corrosion-resistant and pressure-resistant material.

[0012] Further, the corrosion-resistant and pressure-resistant material is titanium or titanium alloy.

[0013] Further, the gas inlet and the gas outlet of the gas storage tank are connected with the seventh pipeline and the gas inlet pipeline respectively through NPT1 / 8-6 sleeve joint.

[0014] Further, the gas inlet and the gas outlet of the air pump are connected with the sixth pipeline through G1 / 8-6 sleeve joint respectively.

[0015] Further, the pressure gauge buffer pipeline is connected with a two-way valve.

[0016] Further, the three-way valve and the two-way valve are sleeve valves.

[0017] Further, the sleeve right-angle valve is a double-head valve, and the front and rear sleeves inside the end far from the second pipeline are replaced with sealing spacers.

[0018] Further, the bottom ends of the liquid sampling pipeline and the gas inlet pipeline are connected with hoses capable of extending into the culture reactor.

[0019] Further, the bottom of the culture reactor is further provided with a magnetic stirrer.

[0020] Compared with the prior art, the utility model has the following technical effects:

[0021] On one hand, the system introduces a gas pump and a precise positive and negative pressure gauge, the gas pump can be used to promote the circulation of gas in the culture system, under the condition of pre-measuring the internal volume of the gas path, the gas content in the system can be accurately calculated according to the reading of the pressure gauge; on the other hand, by setting the liquid sampling pipeline, the gas sampling pipeline and the vacuum sampling pipeline connected with corresponding valves, the circulating gas sample and the liquid sample collection can be considered, which is convenient for real-time analysis of sample components in the culture system; in addition, the gas sampling pipeline is connected with the general gas replacement system, so that the replacement of the gas in the culture system can be realized; it can be seen that the utility model realizes the sampling in the process of purging, circulating and running, solves the problem that the existing anaerobic culture system cannot consider the gas phase and liquid phase sample collection and gas circulation, and is more beneficial to the enrichment culture of microorganisms under absolute anaerobic or anaerobic conditions; in addition, the utility model has the advantages of simple structure and stability, can provide an environment for collecting data accurately and calculating related data for the microbial culture process involving gas metabolism, and can improve the gas path according to actual needs, so that the culture of different microorganisms is applicable, and the utility model has strong applicability.

[0022] The utility model discloses a rigid metal pipeline and a metal gas storage tank are introduced, the headspace of the system is increased, and the air tightness of the system is ensured, and the system can better resist pressure than a general system connected with a flexible pipeline and an aluminum / plastic gas bag.Moreover, compared with silica gel pipes or other plastic pipes used in the existing culture system, the service life is greatly improved, and the sealing performance is better.At the same time, the culture reactor and the gas storage tank are made of pressure-resistant and corrosion-resistant materials, and the materials themselves are not light-transmitting, so that the problem of oxygen production of photosynthetic microorganisms caused by the use of Hungate bottles and Schlenk bottles in the existing culture system is completely avoided.In summary, the utility model is more suitable for the culture of absolute anaerobic microorganisms.

[0023] The utility model discloses a clamp sleeve valve, the air tightness of the connection between the pipelines is higher, air infiltration is effectively prevented, not only beneficial to the enrichment culture of microorganisms under absolute anaerobic or anaerobic conditions, but also make the pressure resistance range of the whole culture system reach-0.3MPa-50MPa, convenient for the switching of negative pressure and positive pressure, and also beneficial to the culture under various pressure conditions in the simulation of natural environment. ACCURACY

[0024] Figure 1 It is a structural schematic diagram of the utility model;

[0025] Figure 2 It is a gas purging mode schematic diagram of the utility model;

[0026] Figure 3 It is a gas replacement mode schematic diagram of the utility model;

[0027] Figure 4The gas circulation mode schematic diagram of the utility model;

[0028] In the figure: 1, liquid sampling pipeline; 2, gas outlet pipeline; 3, gas inlet pipeline; 4, first pipeline; 5, second pipeline; 6, third pipeline; 7, fourth pipeline; 8, fifth pipeline; 9, sixth pipeline; 10, seventh pipeline; 11, vacuum sampling pipeline. DETAILED DESCRIPTION

[0029] The specific content of the utility model is further explained and described in detail in connection with the following embodiments.

[0030] As Figure 1 shown, an absolute anaerobic gas-producing or gas-consuming microorganism culture system includes a culture reactor, a gas storage tank and a gas pump, three through holes are formed at the top of the culture reactor, the three through holes are respectively connected with a liquid sampling pipeline 1, a gas outlet pipeline 2 and a gas inlet pipeline 3, and the liquid sampling pipeline 1 and the gas inlet pipeline 3 are both connected with a two-way valve;

[0031] The gas outlet pipeline 2 is connected with a first pipeline 4 and a second pipeline 5 through a three-way valve, the first pipeline 4 is connected with a third pipeline 6 and a fourth pipeline 7 through a three-way valve, the fourth pipeline 7 is connected with a pressure gauge buffer pipeline through a three-way, the pressure gauge buffer pipeline is connected with a pressure gauge, and the pressure gauge buffer pipeline is connected with a two-way valve;

[0032] The second pipeline 5 is connected with a sleeve right-angle valve, the sleeve right-angle valve is a double-head valve, and the front and rear sleeves inside the end away from the second pipeline 5 are replaced with a sealing spacer, that is, as Figure 1 shown, the front and rear sleeves inside the vertically upward end are replaced with a sealing spacer, so that the second pipeline 5 is isolated from the outside air, and when the sleeve right-angle valve is opened, the gas inside the second pipeline 5 and the gas outlet pipeline 2 can be prevented from exchanging with the outside air, and then the circulating gas is collected by using a gas phase sampling needle to pierce the spacer, and the circulating gas is analyzed and tested by using a gas chromatography method, so as to determine the gas composition in the culture system;

[0033] When it is necessary to exhaust the sleeve right-angle valve, only the nut at the upward end of the sleeve right-angle valve is loosened, so that the sealing spacer is separated from the port of the sleeve right-angle valve, and then the gas can be exhausted;

[0034] The gas inlet pipeline 3 is connected with the gas outlet of the gas storage tank, the gas inlet of the gas storage tank is connected with a seventh pipeline 10, the gas storage tank is connected with a vacuum sampling pipeline 11, the vacuum sampling pipeline 11 includes a vertical pipe segment, a first horizontal pipe segment and a second horizontal pipe segment connected through a three-way valve, the vertical pipe segment is connected with a vacuum valve, and the second horizontal pipe segment is connected with a two-way valve;

[0035] When it is needed to vacuumize the inside of the culture system, the vacuum valve is connected with the vacuum pump; when it is needed to collect gas, the vacuum valve is connected with the sampling tube first, the vacuum pump is connected with the second horizontal pipe section, the air inside the sampling tube, the vertical pipe section and the second horizontal pipe section is extracted by the vacuum pump, then the two-way valve connected with the second horizontal pipe section is closed, the vertical pipe section is connected with the first horizontal pipe section through the three-way valve, and the circulating gas is collected for testing;

[0036] The air inlet and air outlet of the air pump are respectively connected with the sixth pipeline 9, the sixth pipeline 9 connected with the air inlet of the air pump is connected with the fourth pipeline 7 and the fifth pipeline 8 through a three-way valve, and the sixth pipeline 9 connected with the air outlet of the air pump is connected with the seventh pipeline 10 and the fifth pipeline 8 through a three-way valve.

[0037] Preferably, the culture reactor, the gas storage tank, the liquid sampling pipeline 1, the air inlet pipeline 3, the air outlet pipeline 2, the first pipeline 4, the second pipeline 5, the third pipeline 6, the fourth pipeline 7, the fifth pipeline 8, the sixth pipeline 9, the seventh pipeline 10, the pressure gauge buffer pipeline and the vacuum sampling pipeline 11, and the three-way valve and the two-way valve are all made of corrosion-resistant and pressure-resistant materials.

[0038] Preferably, the corrosion-resistant and pressure-resistant materials are titanium or titanium alloy, and in general cases, when the corrosion of the culture solution is small, the corrosion-resistant and pressure-resistant materials can also be stainless steel.

[0039] Preferably, the air inlet and air outlet of the gas storage tank are respectively connected with the seventh pipeline 10 and the air inlet pipeline 3 through NPT1 / 8-6 sleeve joints.

[0040] Preferably, the air inlet and air outlet of the air pump are respectively connected with the sixth pipeline 9 through G1 / 8-6 sleeve joints.

[0041] In actual application, other corrosion-resistant joints can also be used instead of the NPT1 / 8-6 sleeve joints and the G1 / 8-6 sleeve joints.

[0042] Preferably, the three-way valve and the two-way valve are both sleeve valves, and the air tightness between the pipelines is higher, so that the system can withstand a pressure of-0.3MPa to 50MPa, and various gas-producing or gas-consuming microorganism cultures can be carried out in a pressure environment.

[0043] Preferably, the bottom ends of the liquid sampling pipeline 1 and the air inlet pipeline 3 are both connected with hoses that can extend into the culture reactor, the hose connected with the bottom end of the liquid sampling pipeline 1 is conducive to collecting liquid samples from the inside of the culture reactor, and the hose connected with the bottom end of the air inlet pipeline 3 can promote the transfer of gas between gas and liquid when air is introduced.

[0044] Preferably, a magnetic stirrer is further arranged at the bottom of the culture reactor, for promoting the mass transfer in the microorganism reactor.

[0045] Preferably, the volume of the gas storage tank 3 is 3L, but other sizes of gas storage tanks can also be selected according to the actual situation.

[0046] The working principle of this utility model is as follows:

[0047] 1) such as Figure 2 As shown, the purging mode is as follows: Connect the gas cylinder to the third pipe 6, and connect the third pipe 6 and the fourth pipe 7, the fourth pipe 7 and the sixth pipe 9, the sixth pipe 9 and the seventh pipe 10, and the second pipe 5 and the outlet pipe 2 through the corresponding three-way valves. Open the two-way valve connected to the inlet pipe 3 and the clamping right-angle valve connected to the second pipe 5. At the same time, loosen the nut at the top of the clamping right-angle valve. The gas inside the gas cylinder enters the culture reactor through the third pipe 6, the fourth pipe 7, the sixth pipe 9, the seventh pipe 10, the gas storage tank and the inlet pipe 3 in sequence, and then flows out from the culture reactor, flowing out through the outlet pipe 2 and the second pipe 5 in sequence, thus completing the purging of the gas inside the microbial culture system.

[0048] 2) such as Figure 3 As shown, the gas replacement mode is as follows: First, connect the vacuum pump to the vacuum valve. Through the three-way valve, connect the outlet pipe 2 to the first pipe 4, the first pipe 4 to the fourth pipe 7, the fourth pipe 7 to the fifth pipe 8, and the fifth pipe 8 to the seventh pipe 10. Open the two-way valve connected to the inlet pipe 3. At the same time, connect the vertical section and the first horizontal section of the vacuum sampling pipe 11 to facilitate the vacuum pump to extract gas from the gas storage tank, the seventh pipe 10, the fifth pipe 8, the fourth pipe 7, the first pipe 4, the outlet pipe 2, and the culture reactor. Then, turn off the vacuum pump and switch the gas replacement device valve to the gas cylinder containing the required gas. Connect the gas cylinder to the vacuum valve and blow inert gas into the gas storage tank, the seventh pipe 10, the fifth pipe 8, the fourth pipe 7, the first pipe 4, the outlet pipe 2, and the culture reactor. Repeat this pumping and blowing cycle 3 to 5 times to complete the gas replacement inside the microbial culture system.

[0049] In addition, when it is only necessary to replace the gas inside the gas storage tank, the seventh pipe 10, the fifth pipe 8, the fourth pipe 7 and the first pipe 4, the gas outlet pipe 2 and the first pipe 4 are disconnected through the three-way valve, and the two-way valve connected to the gas inlet pipe 3 is closed. The gas replacement device is used to repeat the above-mentioned pumping and blowing cycle 3 to 5 times to complete the gas replacement in the pipeline.

[0050] 3) such as Figure 4As shown, the gas circulation mode: through the corresponding three-way valve, connecting the outlet pipe 2 and the first pipe 4, connecting the first pipe 4 and the fourth pipe 7, connecting the fourth pipe 7 and the sixth pipe 9, the sixth pipe 9 and the seventh pipe 10, and opening the two-way valve connected with the inlet pipe 3, the circulating gas flows out from the culture reactor, and then passes through the outlet pipe 2, the first pipe 4, the fourth pipe 7, the sixth pipe 9, the gas pump, the seventh pipe 10, the gas storage tank and the inlet pipe 3 in turn and returns to the culture reactor;

[0051] In addition, the process of collecting gas phase samples and liquid phase samples is as follows:

[0052] When it is necessary to collect circulating gas, the outlet pipe 2 and the second pipe 5 are connected through the three-way valve, the straight angle valve connected with the second pipe 5 is opened, the gas phase sampling needle is aimed at one end of the straight angle valve, and the circulating gas of the culture system is extracted by piercing the septum; or, the vacuum valve is connected with the sampling pipe, the vacuum pump is connected with the second horizontal pipe section of the vacuum sampling pipe 11, the vertical pipe section of the vacuum sampling pipe 11 is connected with the second horizontal pipe section through the three-way valve, and the air in the sampling pipe, the vertical pipe section and the second horizontal pipe section is extracted by the vacuum pump, then the two-way valve connected with the second horizontal pipe section is closed, the vertical pipe section and the first horizontal pipe section are connected through the three-way valve, and the circulating gas is collected into the sampling pipe;

[0053] When it is necessary to collect the culture liquid, the two-way valve connected with the liquid sampling pipe 1 is opened, and the liquid sample is extracted from the sampling pipe 1 through the needle tube.

[0054] The microbial culture system provided in the embodiment considers gas replacement, gas phase sample and liquid phase sample collection, and positive and negative pressure culture, and provides strong support for simulating microbial growth in various positive or negative pressure natural environments.

Claims

1. A culture system for absolutely anaerobic gas-producing or gas-consuming microorganisms, characterized in that, It includes a culture reactor, a gas storage tank and a gas pump. The top of the culture reactor is connected to a liquid sampling pipe (1), an outlet pipe (2) and an inlet pipe (3), and both the liquid sampling pipe (1) and the inlet pipe (3) are connected to two-way valves. The air outlet pipe (2) is connected to the first pipe (4) and the second pipe (5) through a three-way valve. The first pipe (4) is connected to the third pipe (6) and the fourth pipe (7) through a three-way valve. The fourth pipe (7) is connected to a pressure gauge buffer pipe through a three-way valve. The pressure gauge buffer pipe is connected to the pressure gauge. The second pipe (5) is connected to a compression fitting right-angle valve. The air inlet pipe (3) is connected to the air outlet of the gas storage tank. The air inlet of the gas storage tank is connected to the seventh pipe (10). The gas storage tank is connected to the vacuum sampling pipe (11). The vacuum sampling pipe (11) includes a vertical pipe section, a first horizontal pipe section and a second horizontal pipe section connected by a three-way valve. The vertical pipe section is connected to a vacuum valve and the second horizontal pipe section is connected to a two-way valve. The air pump has a sixth pipe (9) connected to its air inlet and air outlet respectively. The sixth pipe (9) connected to the air inlet of the air pump is connected to the fourth pipe (7) and the fifth pipe (8) through a three-way valve. The sixth pipe (9) connected to the air outlet of the air pump is connected to the seventh pipe (10) and the fifth pipe (8) through a three-way valve.

2. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to claim 1, characterized in that, The culture reactor, gas storage tank, liquid sampling pipeline (1), air inlet pipeline (3), air outlet pipeline (2), first pipeline (4), second pipeline (5), third pipeline (6), fourth pipeline (7), fifth pipeline (8), sixth pipeline (9), seventh pipeline (10), pressure gauge buffer pipeline and vacuum sampling pipeline are all made of corrosion-resistant and pressure-resistant materials.

3. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to claim 2, characterized in that, The corrosion-resistant and pressure-resistant material is titanium or a titanium alloy.

4. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, The air inlet and outlet of the gas storage tank are connected to the seventh pipe (10) and the air inlet pipe (3) respectively via NPT1 / 8-6 compression fittings.

5. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, The air pump's inlet and outlet are connected to the sixth pipe (9) via G1 / 8-6 compression fittings.

6. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, The pressure gauge buffer pipe is connected to a two-way valve.

7. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to claim 6, characterized in that, Both the three-way valve and the two-way valve are compression fitting valves.

8. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, The right-angle valve is a double-headed valve, and the front and rear ferrules inside the end away from the second pipe (5) are replaced with sealing gaskets.

9. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, Both the liquid sampling pipe (1) and the air inlet pipe (3) are connected to flexible tubes that can extend into the culture reactor.

10. The absolutely anaerobic gas-producing or gas-consuming microbial culture system according to any one of claims 1 to 3, characterized in that, The bottom of the culture reactor is also equipped with a magnetic stirrer.

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