CN204401000U - A kind of ocean nitrous acid salt form methane anaerobic oxidized microbial cultivation device - Google Patents

Abstract

The utility model discloses a marine nitrite type methane anaerobic oxidation microorganism cultivation device. It includes a reactor system and a controller system, wherein the reactor system includes a reaction tank and a low-temperature water tank; the upper part of the reaction tank is a reaction tank cover, and a stirring shaft is arranged in the middle of the reaction tank cover. The upper end of the stirring shaft is connected to a stirring motor, and the lower end is connected to a stirring There are also liquid sampling pipes, gas sampling ports, air inlet pipes, air outlet pipes, temperature probes, DO probes, pH probes, conductivity probes and air pressure probes on the cover of the reaction tank; the low temperature water tank is mainly composed of compressors, condensers and The circulating pump is composed of a temperature probe and a liquid level probe on the side wall. The utility model can provide the anaerobic, low temperature, high pressure and high salinity environment necessary for the growth of marine nitrite-type methane anaerobic oxidation microorganisms, and maintain the stable and efficient operation of the device through various probes and control systems, thereby Realize the rapid cultivation of marine nitrite anaerobic methane oxidation microorganisms.

Description

A marine nitrite type methane anaerobic oxidation microbial cultivation device

technical field

The utility model relates to a marine nitrite type methane anaerobic oxidation microorganism cultivation device.

Background technique

As an important energy source, methane plays an important role in human production and life. At the same time, methane is the most abundant hydrocarbon in the atmosphere. Its contribution to global warming is second only to CO ₂ . At present, the "contribution rate" to global warming is as high as 15%, and the greenhouse effect it causes is equimolar 20 to 30 times that of CO ₂ . It is reported that 85% of the world's annual methane production and 60% of its consumption are based on the action of microorganisms. Anaerobic oxidation of methane by microorganisms can make most of the methane gas (more than 90%) be consumed in large quantities before entering the atmosphere. Therefore, anaerobic methane oxidation plays a role that cannot be ignored in the global methane emission control process, and it can effectively alleviate the increasingly serious greenhouse effect.

Nitrite-dependent anaerobic methane oxidation (nitrite-dependent anaerobic methane oxidation) is a newly discovered anaerobic oxidation process of methane, and its reaction equation is shown in formula (1). Gene sequences of nitrite-type anaerobic methane-oxidizing microorganisms have been detected in many ecosystems, and this response has been shown to play an important role in methane control in freshwater wetland systems. At present, researchers at home and abroad have obtained a variety of freshwater nitrite-type methane anaerobic oxidation enrichments, and have done a lot of research on freshwater nitrite-type methane anaerobic oxidation microorganisms. However, no marine nitrite-type anaerobic oxidation enrichment has been successfully enriched, although the gene sequences of nitrite-type anaerobic methane oxidation microorganisms have been detected in marine ecosystems.

3CH ₄ +8NO ₂ ^- +8H ⁺ → 3CO ₂ +4N ₂ +10H ₂ O   (1)

(△G ^0' =－928 kJ mol ^-1 CH ₄ )

Due to the harsh growth conditions of marine nitrite-type methane-oxidizing microorganisms, the laboratory needs to provide a stable anaerobic, low-temperature, high-pressure and high-salinity environment to obtain the enrichment of the microorganisms. The cultivation device puts forward very high requirements, and it is necessary to design a stable and efficient marine nitrite-type methane anaerobic oxidation microbial cultivation device.

Contents of the invention

The purpose of the utility model is to provide a marine nitrite-type methane anaerobic oxidation microorganism cultivation device to overcome the disadvantages of the existing cultivation device such as no pressure resistance, instability, poor robustness, poor sealing, and lack of an alarm system.

The marine nitrite-type methane anaerobic oxidation microbial cultivation device includes a reactor system and a controller system. The reactor system includes a reaction tank, a low-temperature water tank, a cover plate of the reaction tank, a stirring motor, a stirring shaft, a stirring paddle, a temperature probe of the reaction tank, Reaction tank DO probe, reaction tank pH probe, reaction tank conductivity probe, head air pressure probe, liquid sampling tube, gas sampling port, air outlet pipe, air outlet valve, inlet pipe, inlet valve, compressor, condensation pipe, tank liquid Position probe, water tank temperature probe, water tank circulation pump, water tank water inlet, cork, water tank drain pipe, water tank drain valve, sampling needle; There is a stirring shaft in the middle of the cover plate, the upper end of the stirring shaft is connected to the stirring motor, and the lower end is connected to the stirring paddle. The reaction tank cover is also equipped with a reaction tank temperature probe, a reaction tank DO probe, a reaction tank pH probe, a reaction tank conductivity probe, and a top Air pressure probe, liquid sampling pipe, gas sampling port, air outlet pipe and air inlet pipe, among which air outlet valve and air inlet valve are distributed on the air outlet pipe and air inlet pipe; the upper end of the low temperature water tank is provided with a water tank water inlet, and plugged with a cork, The lower left end of the low-temperature water tank is provided with a water tank drain pipe, and a water tank drain valve is arranged on it. The right side of the low-temperature water tank is respectively equipped with a compressor, a water tank liquid level probe, a water tank temperature probe and a condensation pipe from top to bottom, and a water tank is arranged at the lower part of the low-temperature water tank The circulating pump; the controller system is connected with the temperature probe of the reaction tank, the DO probe of the reaction tank, the pH probe of the reaction tank, the conductivity probe of the reaction tank, the head air pressure probe, the water tank liquid level probe and the water tank temperature probe.

The upper part of the reaction tank is cylindrical, the lower part is hemispherical, the ratio of height to diameter of the upper cylindrical shape is 1-2:1, and the diameter of the lower hemispherical shape is equal to the diameter of the upper cylindrical shape. The volume ratio of the reaction tank to the low-temperature water tank is 0.2-0.3:1, and the distance between the bottom of the reaction tank and the bottom of the low-temperature water tank is 5-10 cm. The ratio of the diameter of the stirring paddle to the diameter of the reaction tank is 0.3-0.5:1, and the ratio of the distance from the cover plate of the reaction tank to the total height of the reaction tank is 0.7-0.8:1. The upper end of the liquid sampling tube is sealed by a rubber plug, and the lower end is inserted into the liquid surface by 1 to 3 cm; the gas sampling port is also sealed by a rubber plug; the sampling needle takes a liquid sample through the liquid sampling tube, and a gas sample through the gas sampling port. The sampling port adds acid and alkali liquid to the reaction tank for manual adjustment of pH. The lower end of the inlet pipe is a 45° slope, the slope faces the stirring shaft, and the ratio of the distance from the lower end to the reaction tank cover to the total height of the reaction tank is 0.6-0.7:1.

Compared with the prior art, the utility model has beneficial effects: 1) The growth conditions of marine nitrite-type methane anaerobic oxidation microorganisms are very harsh, and the utility model provides them with anaerobic, low temperature, high pressure and high salinity cultivation environment, and monitor these environmental indicators through various probes; 2) Marine nitrite-type methane-oxidizing microorganisms are very slow-growing microorganisms, and their enrichment and cultivation is a long process, which lasts for several years or even ten years , so it is necessary to ensure the long-term stable operation of the cultivation device. The utility model provides a complete control and alarm system. When the signals monitored by various probes exceed the appropriate range, an alarm will be issued immediately or the power supply will be cut off to reduce the damage caused by instrument failure or operation errors. loss; 3) In order to reduce the probability of device failure due to the failure of individual probes, the utility model adopts a method of monitoring the same index with multiple probes, such as: if the temperature is too high, the temperature probe in the tank, the temperature probe in the tank and the air pressure If the temperature is too low or even freezes, the temperature probe in the tank, the temperature probe in the tank, the air pressure probe and the conductivity probe (after part of the water freezes, the conductivity of the remaining water will increase rapidly) will give an alarm. If one probe fails, the other probes can continue to work independently. 4) The utility model has a smart design and a compact device. The openings of all reaction tanks are set on the cover plate of the reaction tank, which eliminates the possibility of culture medium leakage caused by poor sealing of the openings and increases the ease of use of the device. In addition, The special design of the liquid sampling tube facilitates the collection of liquid samples and increases the integrity of the device.

Description of drawings

Figure 1 is a schematic diagram of the structure of a marine nitrite-type methane anaerobic oxidation microbial culture device.

Detailed ways

As shown in Figure 1, the marine nitrite-type methane anaerobic oxidation microbial cultivation device includes a reactor system 1, a controller system 2, and the reactor system 1 includes a reaction tank 3, a low-temperature water tank 4, a reaction tank cover plate 5, and a stirring motor 6. Stirring shaft 7, stirring paddle 8, reaction tank temperature probe 9, reaction tank DO probe 10, reaction tank pH probe 11, reaction tank conductivity probe 12, head air pressure probe 13, liquid sampling tube 14, gas sampling port 15 , air outlet pipe 16, air outlet valve 17, air inlet pipe 18, air inlet valve 19, compressor 20, condensation pipe 21, water tank liquid level probe 22, water tank temperature probe 23, water tank circulation pump 24, water tank water inlet 25, cork 26 , water tank drain pipe 27, water tank drain valve 28, sampling needle 29; Reaction tank 3 is immersed in the water bath of low-temperature water tank 4, reaction tank 3 upper end is reaction tank cover plate 5, is provided with stirring shaft 7 in the middle of reaction tank cover plate 5, The upper end of the stirring shaft 7 is connected to the stirring motor 6, and the lower end is connected to the stirring paddle 8. The reaction tank cover plate 5 is also provided with a reaction tank temperature probe 9, a reaction tank DO probe 10, a reaction tank pH probe 11, a reaction tank conductivity probe 12, a top Air pressure probe 13, liquid sampling pipe 14, gas sampling port 15, outlet pipe 16 and inlet pipe 18, wherein outlet valve 17 and inlet valve 19 are distributed on the outlet pipe 16 and inlet pipe 18; The water tank inlet 25 is plugged with a cork 26. The lower left end of the low-temperature water tank 4 is provided with a water tank drain pipe 27 on which a water tank drain valve 28 is provided. The right side of the low-temperature water tank 4 is respectively provided with a compressor 20 and a water tank from top to bottom. Liquid level probe 22, water tank temperature probe 23 and condensation pipe 21, the lower part of the low temperature water tank 4 is provided with a water tank circulation pump 24; controller system 2 and reaction tank temperature probe 9, reaction tank DO probe 10, reaction tank pH probe 11, reaction tank The conductivity probe 12, the head air pressure probe 13, the tank liquid level probe 22 and the tank temperature probe 23 are connected.

The upper part of the reaction tank 3 is cylindrical, the lower part is hemispherical, the ratio of height to diameter of the upper cylindrical shape is 1-2:1, and the diameter of the lower hemispherical shape is equal to the diameter of the upper cylindrical shape. The volume ratio of the reaction tank 3 to the low-temperature water tank 4 is 0.2-0.3:1, and the distance between the bottom of the reaction tank 3 and the bottom of the low-temperature water tank 4 is 5-10 cm. The ratio of the diameter of the stirring paddle 8 to the diameter of the reaction tank 3 is 0.3-0.5:1, and the ratio of the distance from the cover plate 5 of the reaction tank to the total height of the reaction tank 3 is 0.7-0.8:1. The upper end of the liquid sampling tube 14 is sealed by a rubber plug, and the lower end is inserted into the liquid surface by 1 to 3 cm; the gas sampling port 15 is also sealed by a rubber plug; Samples are also added to the reaction tank 3 through the gas sampling port 15 to manually adjust the pH. The lower end of the air inlet pipe 18 is a 45° slope, the slope faces the stirring shaft 7, and the ratio of the distance from the lower end to the reaction tank cover plate 5 to the total height of the reaction tank 3 is 0.6-0.7:1.

The marine nitrite-type methane anaerobic oxidation microorganism culture method is: inoculate the marine sediment with a volume of 30-40% of the volume of the reaction tank 3 in the reaction tank 3, and add artificial seawater culture with a volume of 30-40% of the volume of the reaction tank 3 Base, so that the lower ends of the reaction tank temperature probe 9, reaction tank DO probe 10, reaction tank pH probe (11), reaction tank conductivity probe 12, liquid sampling tube 14 and air inlet tube 18 are not below the liquid surface, and the stirring motor 6 is turned on , the stirring motor 6 drives the stirring paddle 8 to rotate through the stirring shaft 7. Under the impetus of the stirring paddle 8, the liquid in the center of the reaction tank 3 flows from top to bottom, and the surrounding liquid flows from bottom to top, and the outlet valve 17 and the inlet valve are opened. 19. Pass methane gas into the reaction tank 3 through the inlet pipe 18, and the excess gas leaves the reaction tank 3 from the gas outlet pipe 16. After the DO probe 10 reading of the reaction tank is lower than 0.2mg/L, first close the gas outlet valve 17, and wait for After the reading of the head air pressure probe 13 is higher than 400 kPa, close the intake valve 19 immediately, and make the reading of the head air pressure probe 13 not higher than 500 kPa. If the reading of the reaction tank pH probe 11 is lower than 7.0 or higher than 8.0, Add an appropriate amount of NaOH or HCl to the reaction tank 3 through the gas sampling port 15 to adjust the pH to about 7.5; add distilled water to the low-temperature water tank 4 until the liquid level is 0.8 of the total internal height of the low-temperature water tank 4, and set the temperature of the low-temperature water tank 4 After stabilization, the readings of the reaction tank temperature probe 9 and the water tank temperature probe 23 should be in the range of 8 to 12 °C; the controller system 2 monitors the reaction tank 3 and the low temperature through the reaction tank temperature probe 9 and the water tank temperature probe 23 respectively. When the temperature in the water tank 4 exceeds the allowable range of 8-12°C, the power supply of the low-temperature water tank 4 is cut off and an alarm is issued. The controller system 2 monitors the dissolved oxygen in the reaction tank 3 through the DO probe 10 of the reaction tank. When the reading When it is greater than 0.2mg/L, cut off the power supply of the low-temperature water tank 4 and send out an alarm. The controller system 2 monitors the conductivity of the liquid in the reaction tank 3 through the conductivity probe 12 of the reaction tank. When the conductivity value exceeds ±20% of the initial value , cut off the power supply of the low temperature water tank 4, and send an alarm, the controller system 2 monitors the gas pressure in the reaction tank 3 through the head air pressure probe 13, when the air pressure is greater than 600kPa or less than 200kPa, cut off the power supply of the low temperature water tank 4, and send an alarm, The controller system 2 monitors the pH value of the liquid in the reaction tank 3 through the pH probe 11 of the reaction tank. When the pH value exceeds the normal range of 7.0 to 8.0, an alarm is issued, waiting for manual adjustment. The controller system 2 monitors through the water tank liquid level probe 22 The liquid level height in the low temperature water tank 4, when the ratio of the liquid level height to the total internal height of the low temperature water tank 4 is greater than 0.9 or less than 0.7, an alarm is issued and manual adjustment is awaited.

Claims (6)

1. an ocean nitrous acid salt form methane anaerobic oxidized microbial cultivation device, it is characterized in that comprising reactor assembly (1), controller system (2), reactor assembly (1) comprises retort (3), low temperature tank (4), retort cover plate (5), agitator motor (6), stir shaft (7), stirring rake (8), retort temp probe (9), retort DO pops one's head in (10), retort pH pops one's head in (11), retort conductivity probe (12), head space air pressure probe (13), liquid sampling pipe (14), gas sampling mouth (15), escape pipe (16), air outlet valve (17), inlet pipe (18), intake valve (19), compressor (20), prolong (21), water level probe (22), tank temp probe (23), aquarium circulating pump (24), tank water-in (25), cork stopper (26), sink drain (27), drain valve of wash tank (28), sampling probe (29), retort (3) is immersed in the water-bath of low temperature tank (4), retort (3) upper end is retort cover plate (5), stir shaft (7) is provided with in the middle of retort cover plate (5), stir shaft (7) upper end connects agitator motor (6), lower end connects stirring rake (8), retort cover plate (5) is also provided with retort temp probe (9), retort DO pops one's head in (10), retort pH pops one's head in (11), retort conductivity probe (12), head space air pressure probe (13), liquid sampling pipe (14), gas sampling mouth (15), escape pipe (16) and inlet pipe (18), wherein escape pipe (16) and inlet pipe (18) are distributed with air outlet valve (17) and intake valve (19), low temperature tank (4) upper end is provided with tank water-in (25), and be plugged with cork stopper (26), low temperature tank (4) lower-left end is provided with sink drain (27), which is provided with drain valve of wash tank (28), low temperature tank (4) right side is respectively equipped with compressor (20), water level probe (22), tank temp probe (23) and prolong (21) from top to bottom, and low temperature tank (4) bottom is provided with aquarium circulating pump (24), controller system (2) is popped one's head in (10) with retort temp probe (9), retort DO, retort pH pops one's head in (11), retort conductivity probe (12), head space air pressure are popped one's head in (13), water level is popped one's head in, and (22) are connected with tank temp probe (23).

2. a kind of ocean according to claim 1 nitrous acid salt form methane anaerobic oxidized microbial cultivation device, it is characterized in that: described retort (3) top is cylindrical, bottom is semisphere, Upper cylindrical shape aspect ratio is 1 ~ 2:1, lower hemispherical diameter and Upper cylindrical shape equal diameters.

3. a kind of ocean according to claim 1 nitrous acid salt form methane anaerobic oxidized microbial cultivation device, it is characterized in that: described retort (3) is 0.2 ~ 0.3:1 with the volume ratio of low temperature tank (4), retort (3) bottom is 5 ~ 10cm with the distance of low temperature tank (4) bottom.

4. a kind of ocean according to claim 1 nitrous acid salt form methane anaerobic oxidized microbial cultivation device, it is characterized in that: described stirring rake (8) diameter and the ratio of retort (3) diameter are 0.3 ~ 0.5:1, the ratio from the distance of retort cover plate (5) and retort (3) total height is 0.7 ~ 0.8:1.

5. a kind of ocean according to claim 1 nitrous acid salt form methane anaerobic oxidized microbial cultivation device, is characterized in that: described liquid sampling pipe (14) upper end is by rubber stopper seal, and liquid level 1 ~ 3cm is inserted in lower end; Gas sampling mouth (15) is also by rubber stopper seal; Sampling probe (29) gets liquid sample by liquid sampling pipe (14), gets gaseous sample by gas sampling mouth (15), also in retort (3), adds acid & alkali liquid by gas sampling mouth (15), carries out the manual regulation of pH.

6. a kind of ocean according to claim 1 nitrous acid salt form methane anaerobic oxidized microbial cultivation device, it is characterized in that: described inlet pipe (18) lower end is 45 ° of inclined-planes, inclined-plane is towards stir shaft (7), and lower end is 0.6 ~ 0.7:1 from the ratio of the distance of retort cover plate (5) and retort (3) total height.