CN109022258B

CN109022258B - Archaea culture device suitable for growth in low-temperature micro-aerobic environment

Info

Publication number: CN109022258B
Application number: CN201811063231.1A
Authority: CN
Inventors: 臧淑英; 谢桂林; 李苗; 解瑞峰; 马欣然; 智刚; 张鹏飞; 程进; 陈伟; 崔薇; 马大龙
Original assignee: Harbin Normal University
Current assignee: Harbin Normal University
Priority date: 2018-09-12
Filing date: 2018-09-12
Publication date: 2022-06-10
Anticipated expiration: 2038-09-12
Also published as: CN109022258A

Abstract

Be suitable for archaea culture apparatus of low temperature little oxygen environment growth, belong to microbiology technical field, constitute its characterized in that by adjustable little oxygen culture box (22), constant temperature illumination incubator (19) and aspiration pump (26): adjustable little oxygen culture box (22) including flourishing water box (1), accuse oxygen water (2), transfer trachea (3), scale limit oxygen section of thick bamboo (4), foam buoyancy piece (5), culture dish (6), transfer trachea skin stifled (7), flourishing water box wall (8), flourishing water box chamber (9) and little oxygen space (10), hold in flourishing water box (1) accuse oxygen water (2), during scale limit oxygen section of thick bamboo (4) opening inserts accuse oxygen water (2) down, form little oxygen space (10) bottom scale limit oxygen section of thick bamboo (4), culture dish (6) of adhesion on foam buoyancy piece (5) suspend at the upper surface of accuse oxygen water (2). The archaebacteria culture device suitable for growth in a low-temperature micro-aerobic environment is simple to manufacture, strong in operability, low in cost and obvious in effect.

Description

Archaea culture device suitable for growth in low-temperature micro-aerobic environment

Technical Field

The invention relates to an archaea culture device suitable for growth in a low-temperature micro-aerobic environment, and belongs to the technical field of microbiology.

Background

Archaea were proposed in 1977 by Carl Woese and George Fox because they differ significantly from other prokaryotes in the phylogenetic tree of 16 SrRNA. Archaea is a very special bacterium, mostly lives in extreme environments such as hot spring water, anoxic lake bottom, salt water lake and the like, some lives in extremely high temperature such as intermittent spring or seabed black chimney, some lives in very cold environment or high-salt, strong-acid or strong-alkaline water, some is neutrophilic, can be found in marsh, waste water and soil, and many archaea producing methane live in animal digestive tracts. Archaebacteria have a single diameter of 0.1 to 15 microns, some species form cell clusters or fibers, have a length of 200 microns, are various in shape, have a spherical, rod, spiral, leaf or square shape, have an endomembrane and endomembrane system, have some unique biochemical properties, such as the ability of halobacter to produce ATP using light energy, membrane lipids are linked by ether bonds rather than ester bonds, and are also different in nutrition from conventional organisms, such as sulfur oxidation, etc., protein synthesis is initiated with methionine, ribosomes are insensitive to chloramphenicol, RNA polymerase and eukaryotic cells are similar, DNA has introns and binds histones, cellular lipids are not convertible, cell walls do not contain peptidoglycan, some are protein-based and some contain heteropolysaccharides, all do not contain muramic acid, D-type amino acids and diaminopimelic acid. The amount of information inherited by archaea is small. From the RNA evolution tree, archaea fall into two categories: sparomyces perniciae (Crenarchaeota) and eurycotina euryaleota. Archaea have also been classified into three groups: methanogenic bacteria, extreme halophilic bacteria and acidophilic thermophilic bacteria. At present, 22 archaebacteria genomes have been sequenced. Although the number of archaea is sometimes very large in nature and many archaea information can be obtained in high throughput measurement of some environments, it is not known how the physicochemical properties and actions of these archaea are specific, and research on archaea is very small compared with the stock amount and actions thereof because it is difficult to culture archaea by conventional means. Therefore, how to successfully culture the archaebacteria in the laboratory becomes a great problem to be solved urgently, so that by utilizing the water tightness, a culture dish for sample culture is placed in the living environment simulating the archaebacteria to provide micro-oxygen and certain pressure for the archaebacteria, thereby achieving the purpose of successfully culturing the archaebacteria in the laboratory, and the invention is necessary to invent the archaebacteria culture device suitable for growth in the low-temperature micro-oxygen environment.

Disclosure of Invention

In order to overcome the problem that archaea are difficult to culture according to a conventional means, the invention provides an archaea culture device suitable for growth in a low-temperature micro-aerobic environment, the archaea culture device suitable for growth in the low-temperature micro-aerobic environment utilizes the tightness of water, a culture dish for sample culture is placed in an adjustable micro-aerobic culture box simulating the survival environment of the archaea, meanwhile, the adjustable micro-aerobic culture box is placed in a constant-temperature illumination incubator, the low-temperature and illumination conditions are selected, micro-oxygen and certain pressure are provided for the archaea, and therefore the aim of smoothly culturing the archaea in a laboratory is achieved.

The technical scheme adopted by the invention for solving the technical problem is as follows:

the invention relates to an archaebacteria culture device suitable for growth in a low-temperature micro-aerobic environment, which consists of an adjustable micro-aerobic culture box 22, a constant-temperature illumination culture box 19 and an air pump 26, and is characterized in that: the adjustable micro-aerobic culture box 22 is cuboid, the length of the cuboid is 15-20 cm, the width is 15-20 cm, the height is 20-30 cm, the adjustable micro-aerobic culture box 22 comprises a water containing box 1, oxygen control water 2, an air adjusting pipe 3, a scale oxygen limiting cylinder 4, a foam buoyancy block 5, a culture dish 6, an air adjusting pipe leather plug 7, a water containing box wall 8, a water containing box cavity 9 and a micro-aerobic space 10, the oxygen control water 2 is contained in the water containing box 1, the scale oxygen limiting cylinder 4 is inserted into the oxygen control water 2 in the water containing box 1 with an opening facing downwards in the water containing box 1, the micro-aerobic space 10 can be formed at the bottom of the scale oxygen limiting cylinder 4 because the oxygen control water 2 is only partially filled in the scale oxygen limiting cylinder 4, the culture dish 6 adhered on the foam buoyancy block 5 can be suspended on the upper surface of the oxygen control water 2 at the bottom of the micro-aerobic space 10 by virtue of the buoyancy of the foam buoyancy block 5 after bacteria inoculation, one end of the gas regulating pipe 3 is communicated with the micro-oxygen space 10, the other end of the gas regulating pipe is blocked by a gas regulating pipe skin plug 7, and when gas is regulated, the gas regulating pipe skin plug 7 is taken down and communicated with the air extracting pump 26, so that gas in the micro-oxygen space 10 can be extracted, and the content of oxygen in the micro-oxygen space 10 can be regulated; the water containing box 1 is cuboid, the length of the cuboid is 15-20 cm, the width of the cuboid is 15-20 cm, and the height of the cuboid is 20-30 cm; the outer wall of the water containing box 1 is a water containing box wall 8, wherein the water containing box 1 is not provided with an upper bottom wall, the lower bottom and the water containing box walls 8 at the periphery are made of glass or plastics, and the thickness of the glass or plastics is 0.3-0.6 cm; the cavity enclosed by the water containing box wall 8 is a water containing box cavity 9; the oxygen-controlled water 2 is tap water used in daily life; the air adjusting pipe 3 is made of glass or plastic and is U-shaped, the arm length of the U-shaped is 18-28 cm, the cross section of the air adjusting pipe 3 is circular, the outer diameter of the circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, the thickness of the circular ring is 0.1-0.2 cm, and the air adjusting pipe 3 is fixed on the wall of the scale oxygen limiting cylinder 4; the air adjusting pipe leather plug 7 is made of rubber and has elasticity, one end is opened, the other end is closed, the inner diameter is 0.45-0.9 cm, the length is 1-2 cm, the wall thickness is 0.5-1 mm, the air adjusting pipe leather plug is taken down from the air adjusting pipe 3 when the air pump 26 pumps air, and the air adjusting pipe leather plug is sleeved at the outer end of the air adjusting pipe 3 after air pumping; the scale oxygen-limiting cylinder 4 is a container which limits the foam buoyancy block 5 and the culture dish 6 and provides micro-oxygen environment for the cultured archaea in the culture dish 6, is cylindrical, the diameter of the cylinder is 10-16 cm, the height of the cylinder is 18-28 cm, the upper bottom and the peripheral wall of the scale oxygen-limiting cylinder 4 are made of glass or plastic, the thickness of the cylinder is 1-2 mm, the scale oxygen-limiting cylinder 4 is not provided with a lower bottom wall, oxygen-controlling water 2, the foam buoyancy block 5 and the culture dish 6 enter the scale oxygen-limiting cylinder 4, volume scales are arranged on the peripheral wall of the scale oxygen-limiting cylinder 4 from top to bottom, and the volume scales are finished from 0 ml to 1000 ml; the foam buoyancy block 5 is a buoyancy part which is adhered to the lower part of the culture dish 6, provides buoyancy for the culture dish 6 and enables the culture dish 6 to float on the liquid surface of the oxygen control water 2 in the scale oxygen limiting cylinder 4, is made of foam plastic and is in a round cake shape, the diameter of the round cake is 5-12 cm, and the thickness of the round cake is 1-2 cm; the culture dish 6 is a part for culturing archaea, the part poured with the culture medium is a culture dish basal disc, and the part covering the culture dish basal disc is a culture dish cover; the culture medium used in the culture dish 6 is beef extract peptone culture medium or commercially available LB culture medium added with 1-5 ml of living environment leachate to be collected from the sample to be cultured and 1-2 g of sulfur-containing compound; the micro-oxygen space 10 is a closed space formed by filling the oxygen control water 2 into the scale oxygen limiting cylinder 4, the foam buoyancy block 5 and the culture dish 6 are suspended on the upper surface of the oxygen control water 2 in the micro-oxygen space 10, the size of the micro-oxygen space 10 changes along with the change of the air suction amount, the volume scale on the wall of the scale oxygen limiting cylinder 4 can be read in real time, and the pressure in the micro-oxygen space 10 can be controlled by the adding amount and the operation mode of the oxygen control water 2.

The air pump 26 is an independent part for regulating and controlling the size of the micro-oxygen space 10 by pumping gas in the micro-oxygen space 10 through the air regulating pipe 3, and the structure and the function of the air pump 26 are the same as those of a known air pump and comprise an air pump power plug 11, an air pump power line 12, an air pump handle 13, an air pump bracket 14, an air pump air outlet 15, an air pump body 16, a communicating sleeve 17 and an air pump air inlet pipe 18; the structure and the function of the air suction pump power plug 11 are the same as those of a known two-phase power plug, the structure and the function of the air suction pump power cord 12 are the same as those of a known two-strand copper core power cord, the cross section of the copper core is circular, the diameter of the circle is 1-5 mm, the air suction pump handle 13 is made of stainless steel or aluminum alloy, the cross section of the air suction pump handle is circular, the diameter of the circle is 1-3 cm, the length of the circle is 30-40 cm, two ends of the air suction pump handle are bent towards the same direction, the bent length is 5-10 cm, and the two ends of the air suction pump handle are welded on the upper surface of the air suction pump body 16 after being bent, so that a handle shape is formed; the air pump supports 14 are supports welded on the lower surface of the

air pump body

16, 1 air pump is respectively arranged at the front end and the rear end of the air pump body 16, each air pump support 14 is made of stainless steel or aluminum alloy and is solid, the cross section of each air pump support is a trapezoid with a concave upper bottom surface, the length of the upper bottom side of each trapezoid is 3-5 cm, the length of the lower bottom side of each trapezoid is 10-15 cm, and the height of each trapezoid is 3-5 cm; the air outlet 15 of the air pump is an opening through which the micro-oxygen space 10 is extracted by the air pump 26 and then discharged out of the air pump body 16, and is positioned at one end of the air pump body 16, and an iron mesh is arranged at the opening; the air pump body 16 is cylindrical, the diameter of the cylinder is 10-15 cm, the length of the cylinder is 20-30 cm, and the structure and the function of the air pump body 16 are the same as those of a well-known air pump 26; the air intake pipe 18 of the air pump is a long pipe which is positioned at the other end of the air pump body 16 opposite to the air outlet 15 of the air pump and is made of stainless steel or aluminum alloy, the length is 10-20 cm, the cross section of the air intake pipe 18 of the air pump is circular, the outer diameter of the circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, and the thickness of the circular ring is 0.1-0.2 cm; the communicating sleeve 17 is made of rubber and elastic, one end of the communicating sleeve is sleeved at the free end of the air inlet pipe 18 of the air suction pump when the air suction pump 26 sucks air, the other end of the communicating sleeve is sleeved at the outer end of the air adjusting pipe 3, the inner diameter of the communicating sleeve 17 is 0.45-0.9 cm, the length of the communicating sleeve 17 is 10-20 cm, and the thickness of the wall of the communicating sleeve 17 is 0.5-1 mm.

The constant-temperature illumination incubator 19 is cuboid, the length of the cuboid is 50-300 cm, the width of the cuboid is 50-100 cm, and the height of the cuboid is 150-300 cm, and comprises a constant-temperature illumination incubator power plug 20, a constant-temperature illumination incubator power line 21, an adjustable micro-aerobic incubator 22, a light illuminator 23, a transparent support plate 24 and a constant-temperature illumination incubator cavity 25; the structure and the function of the power plug 20 of the constant-temperature illumination incubator are the same as those of a well-known two-phase power plug; the structure and the function of the power line 21 of the constant-temperature illumination incubator are the same as those of a known double-strand copper core power line, the cross section of the copper core is circular, and the diameter of the circle is 1-5 mm; the structure and the function of the illumination lamps 23 are the same as those of the well-known fluorescent lamps, 1 to 4, and the illumination lamps are arranged at the top of the constant-temperature illumination incubator cavity 25; the transparent support plate 24 is made of glass or plastic with high transparency, has the thickness of 0.3-0.8 cm, and is arranged on the support in the two side walls of the constant-temperature illumination incubator 19; the constant temperature illumination incubator cavity 25 is a space inside the constant temperature illumination incubator 19, a transparent support plate 24 is arranged inside, and the adjustable micro-aerobic incubator 22 is placed on the transparent support plate 24.

The invention has the beneficial effects that the archaebacteria culture device suitable for growth in a low-temperature micro-aerobic environment utilizes the tightness of water, a culture dish used for sample culture is placed in an adjustable micro-aerobic culture box simulating the survival environment of the archaebacteria, the adjustable micro-aerobic culture box is placed in a constant-temperature illumination incubator, the low-temperature and illumination conditions are selected, micro-oxygen and certain pressure are provided for the archaebacteria, and the aim of smoothly culturing the archaebacteria in a laboratory is fulfilled. The device for the archaebacteria culture device suitable for growth in the low-temperature micro-aerobic environment is simple to manufacture, strong in operability, low in cost and obvious in effect.

Drawings

The invention is further described below with reference to the accompanying drawings.

FIG. 1 is a schematic diagram of the overall structure of an adjustable micro-aerobic culture box of the archaeal bacteria culture device suitable for growth in a low-temperature micro-aerobic environment.

FIG. 2 is a schematic view of the overall structure of the air pump of the archaeal bacteria culture apparatus suitable for growth in a low-temperature micro-aerobic environment according to the present invention.

FIG. 3 is a schematic view of the overall structure of the constant temperature illumination incubator of the archaeal bacteria culture apparatus suitable for growth in a low temperature micro-aerobic environment of the present invention.

In the figure, 1, a water containing box, 2, oxygen control water, 3, an air adjusting pipe, 4, a scale oxygen limiting cylinder, 5, a foam buoyancy block, 6, a culture dish, 7, an air adjusting pipe skin plug, 8, a water containing box wall, 9, a water containing box cavity, 10, a micro-oxygen space, 11, an air pump power plug, 12, an air pump power cord, 13, an air pump handle, 14, an air pump bracket, 15, an air pump air outlet, 16, an air pump body, 17, a communicating sleeve, 18, an air pump air inlet pipe, 19, a constant-temperature illumination culture box, 20, a constant-temperature illumination culture box power plug, 21, a constant-temperature illumination culture box power cord, 22, an adjustable micro-oxygen culture box, 23, a light, 24, a transparent supporting plate, 25, a constant-temperature illumination culture box cavity and 26, an air pump are arranged.

Detailed Description

The first embodiment is as follows:

as shown in the figure, the archaebacteria culture device suitable for growth in a low-temperature micro-aerobic environment comprises a water containing box 1, oxygen control water 2, an air adjusting pipe 3, a scale oxygen limiting cylinder 4, a foam buoyancy block 5, a culture dish 6, an air adjusting pipe leather plug 7, a water containing box wall 8, a water containing box cavity 9, a micro-aerobic space 10, an air extracting pump power plug 11, an air extracting pump power line 12, an air extracting pump handle 13, an air extracting pump bracket 14, an air extracting pump air outlet 15, an air extracting pump body 16, a communicating sleeve 17, an air extracting pump air inlet pipe 18, a constant-temperature illumination culture box 19, a constant-temperature illumination culture box power plug 20, a constant-temperature illumination culture box power line 21, an adjustable micro-aerobic culture box 22, a light 23, a transparent support plate 24, a constant-temperature illumination culture box cavity 25 and an air extracting pump 26. The adjustable micro-aerobic culture box 22 is a container for providing a micro-aerobic growth environment for archaea, is in a cuboid shape, the length of the cuboid is 15-20 cm, the width of the cuboid is 15-20 cm, the height of the cuboid is 20-30 cm, the adjustable micro-aerobic culture box 22 comprises a water containing box 1, oxygen control water 2, an air adjusting pipe 3, a scale oxygen limiting cylinder 4, a foam buoyancy block 5, a culture dish 6, an air adjusting pipe leather plug 7, a water containing box wall 8, a water containing box cavity 9 and a micro-aerobic space 10, the oxygen control water 2 is contained in the water containing box 1, the scale oxygen limiting cylinder 4 is inserted into the oxygen control water 2 in the water containing box 1 with an opening facing downwards in the water containing box 1, the micro-aerobic space 10 can be formed at the bottom of the scale oxygen limiting cylinder 4 because the oxygen control water 2 is only partially filled in the scale oxygen limiting cylinder 4, the culture dish 6 adhered on the foam buoyancy block 5 can be suspended on the lower bottom of the micro-aerobic space 2 by virtue of the buoyancy block 5 after bacteria inoculation, one end of the gas regulating pipe 3 is communicated with the micro-oxygen space 10, the other end of the gas regulating pipe is plugged by the gas regulating pipe plug 7, and when gas is regulated, the gas regulating pipe plug 7 is taken down and communicated with the air extracting pump 26, so that gas in the micro-oxygen space 10 can be extracted, and the content of oxygen in the micro-oxygen space 10 can be further regulated. The water containing box 1 is a container for containing oxygen control water 2, an air adjusting pipe 3, a scale oxygen limiting cylinder 4, a foam buoyancy block 5 and a culture dish 6, the water containing box 1 is cuboid, the length of the cuboid is 15-20 cm, the width of the cuboid is 15-20 cm, and the height of the cuboid is 20-30 cm; the outer wall of the water containing box 1 is a water containing box wall 8, wherein the water containing box 1 is not provided with an upper bottom wall, the lower bottom and the water containing box walls 8 at the periphery are made of glass or plastics, the thickness of the glass or plastics is 0.3-0.6 cm, and the water containing box walls 8 also form the outer wall of the adjustable micro-aerobic culture box 22; the cavity surrounded by the water containing box wall 8 is a water containing box cavity 9, and the oxygen control water 2 is poured into the water containing box cavity 9. The oxygen control water 2 is tap water used in daily life, and is used for blocking the communication between the inside of the water containing box cavity 9 and outside air after the scale oxygen limiting cylinder 4 is buckled in the water containing box cavity 9 in an inverted mode, so that a closed micro-oxygen space 10 is formed in the scale oxygen limiting cylinder 4, once air is pumped through the air adjusting pipe 3, the pressure in the micro-oxygen space 10 is reduced and is lower than the outside atmospheric pressure, the oxygen control water 2 can enter the scale oxygen limiting cylinder 4 to form a height difference, and therefore the pressure balance is maintained, if the scale oxygen limiting cylinder 4 is placed in the oxygen control water 2, the oxygen control water 2 is continuously added into the water containing box cavity 9, and due to the fact that the pressure balance needs to be maintained inside and outside the scale oxygen limiting cylinder 4, the pressure in the micro-oxygen space 10 can be increased by adding the oxygen control water 2 and is higher than the outside atmospheric pressure, and therefore an environment with adjustable pressure is provided for the growth of archaea, and finally a specific pressure state is maintained. The gas regulating pipe 3 is made of glass or plastic and is U-shaped, the arm length of the U-shaped is 18-28 cm, the cross section of the gas regulating pipe 3 is circular, the outer diameter of a circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, the thickness of the circular ring is 0.1-0.2 cm, the gas regulating pipe 3 is fixed on the wall of the scale oxygen limiting cylinder 4, and when gas is regulated, the gas regulating pipe skin plug 7 is taken down and communicated with the air extracting pump 26, so that gas in the micro-oxygen space 10 can be extracted, and the content of oxygen in the micro-oxygen space 10 can be regulated. The air adjusting pipe leather plug 7 is a plug at the outer end of the air adjusting pipe 3 and is used for plugging the air adjusting pipe 3 after the air pump 26 extracts air in the micro-oxygen space 10 and blocking the communication between the micro-oxygen space 10 and the outside air, the air adjusting pipe leather plug 7 is made of rubber and is elastic, one end of the air adjusting pipe is open, the other end of the air adjusting pipe leather plug is closed, the inner diameter of the air adjusting pipe leather plug is 0.45-0.9 cm, the length of the air adjusting pipe leather plug is 1-2 cm, the wall thickness of the air adjusting pipe leather plug is 0.5-1 mm, the air adjusting pipe leather plug is taken down from the air adjusting pipe 3 when the air pump 26 extracts air, and the air adjusting pipe 3 is sleeved with the extracted air. The scale oxygen-limiting cylinder 4 is a container which limits the foam buoyancy block 5 and the culture dish 6 and provides micro-oxygen environment for the cultured archaea in the culture dish 6, is cylindrical, the diameter of the cylinder is 10-16 cm, the height of the cylinder is 18-28 cm, the upper bottom and the peripheral wall of the scale oxygen-limiting cylinder 4 are made of glass or plastic, the thickness of the cylinder is 1-2 mm, the scale oxygen-limiting cylinder 4 is not provided with a lower bottom wall, and oxygen-controlling water 2, the foam buoyancy block 5 and the culture dish 6 enter the scale oxygen-limiting cylinder 4, the volume scales are arranged on the peripheral wall of the scale oxygen-limiting cylinder 4 from top to bottom, the volume scales begin from 0 ml to end at 1000 ml, and as the glass or the plastic has certain transparency, therefore, after the oxygen control water 2 is injected into the water containing box cavity 9, the liquid level position of the oxygen control water 2 in the scale oxygen limiting cylinder 4 can be seen, and the volume of the micro-oxygen space 10 can be roughly read. The foam buoyancy block 5 is a buoyancy part which is adhered below the culture dish 6 and provides buoyancy for the culture dish 6 to enable the culture dish 6 to float on the liquid surface of the oxygen control water 2 in the scale oxygen limiting cylinder 4, is made of foam plastics and is in a round cake shape, the diameter of the round cake is 5-12 cm, the thickness of the round cake is 1-2 cm, after the internal treatment of the culture dish 6 is completed, namely, a solid culture medium with the thickness of 1-5 mm is poured into a culture dish basal disc, after the solid culture medium is cooled, the culture dish basal disc is placed on a super clean workbench and inoculated with archaebacteria liquid to be cultured, a culture dish cover is covered, a sealing film is wound on the culture dish 6, the culture dish basal disc and the culture dish cover are fixed together, then the culture dish is covered under, the culture dish basal disc is placed on the upper surface of the foam buoyancy block 5 to form an inverted state, the foam buoyancy block 5 and the culture dish 6 are adhered together at the joint of the foam buoyancy block 5 and the culture dish 6 by using a commercially available adhesive, the culture dish 6 is prevented from falling off from the foam buoyancy block 5, and the culture dish 6 is ensured to be firmly positioned on the upper surface of the foam buoyancy block 5 all the time. The culture dish 6 is a part for culturing archaea, the part poured with the culture medium is a culture dish basal disc, and the part covering the culture dish basal disc is a culture dish cover; the culture medium used in the culture dish 6 is beef extract peptone medium or commercial LB medium, such as archaebacteria in frozen soil, added with 1-5 ml of the living environment leachate of the sample to be cultured and 1-2 g of sulfur-containing compounds, and the preparation method of the living environment leachate of the sample collection is as follows: 1 g of frozen soil is taken, 10 ml of water is used for soaking, and a microporous filter membrane is used for filtering, so that the obtained liquid is the living environment leaching liquid of the sample collection place; the preparation method of the inoculated bacterial liquid comprises the following steps: 1 g of frozen soil is taken, 10 ml of water is used for soaking, and after standing for 10 minutes, the supernatant is the inoculated bacterial liquid; when inoculating the bacterium liquid, taking 1-2 microliters of supernatant by using a pipettor, blowing into the surface of the culture medium, taking 5-6 glass beads, covering a culture dish cover, shaking the culture dish for 61-2 minutes, rolling the glass beads back and forth in the shaking process, uniformly coating the archaea to be cultured on the surface of the culture medium, opening the culture dish cover, pouring the glass beads into a bottle containing alcohol, covering the culture dish cover, and winding and fixing the culture dish cover and the culture dish base plate together by using a sealing film to obtain the culture dish 6. The micro-oxygen space 10 is a closed space formed by filling the oxygen control water 2 into the scale oxygen limiting cylinder 4, the foam buoyancy block 5 and the culture dish 6 are suspended on the upper surface of the oxygen control water 2 in the micro-oxygen space 10, the size of the micro-oxygen space 10 changes along with the change of the air suction amount, the volume scale on the wall of the scale oxygen limiting cylinder 4 can be read in real time, and the pressure in the micro-oxygen space 10 can be controlled by the adding amount and the operation mode of the oxygen control water 2. The air pump 26 is an independent part for adjusting and controlling the size of the micro-oxygen space 10 by extracting gas in the micro-oxygen space 10 through the air adjusting pipe 3, the air pump 26 is only used when the adjustable micro-oxygen culture box 22 is arranged, after the adjustable micro-oxygen culture box 22 is arranged, the air pump 26 is stored for next use, and the structure and the function of the air pump 26 are the same as those of a well-known air pump, and the air pump 26 consists of an air pump power plug 11, an air pump power line 12, an air pump handle 13, an air pump support 14, an air pump air outlet 15, an air pump body 16, a communication sleeve 17 and an air pump air inlet pipe 18. The suction pump power plug 11 is structurally and functionally similar to a known two-phase power plug, and can introduce electric energy into the suction pump power cord 12 from the outside. The structure and the function of the air pump power cord 12 are the same as those of a well-known double-strand copper core power cord, the cross section of the copper core is circular, the diameter of the circular copper core is 1-5 mm, and electric energy can be led into an air pump body 16 from an air pump power plug 11. The air pump handle 13 is a component which is convenient for lifting the air pump 26, is made of stainless steel or aluminum alloy, has a circular cross section, has a diameter of 1-3 cm and a length of 30-40 cm, is bent at two ends in the same direction, has a bent length of 5-10 cm, and is welded at two ends on the upper surface of the air pump body 16 after being bent, so that a handle shape is formed. The air extracting pump supports 14 are supports welded on the lower surface of the air extracting pump body 16 and used for supporting and fixing the air extracting pump body 16, the front end and the rear end of the air extracting pump body 16 are respectively provided with 1, each air extracting pump support 14 is made of stainless steel or aluminum alloy and is solid, the cross section of each air extracting pump support is a trapezoid with a concave upper bottom surface, the length of the upper bottom edge of the trapezoid is 3-5 cm, the length of the lower bottom edge of the trapezoid is 10-15 cm, and the height of the trapezoid is 3-5 cm. The air outlet 15 of the air pump is an opening through which the micro-oxygen space 10 is extracted by the air pump 26 and then discharged out of the air pump body 16, and is located at one end of the air pump body 16, and an iron mesh is arranged at the opening and used for preventing dust from entering the air pump 26. The air pump body 16 is cylindrical, the diameter of the cylinder is 10-15 cm, the length of the cylinder is 20-30 cm, the structure and the function of the air pump body 16 are the same as those of a well-known air pump 26, and an inner motor works under the power-on state to drive an impeller to rotate, so that the suction force is formed, and the micro-oxygen space 10 is extracted. The air intake pipe 18 of the air pump is a long pipe which is arranged at the other end of the air pump body 16 opposite to the air outlet 15 of the air pump and is made of stainless steel or aluminum alloy, the length of the air intake pipe is 10-20 cm, the cross section of the air intake pipe 18 of the air pump is circular, the outer diameter of the circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, and the thickness of the circular ring is 0.1-0.2 cm. The communicating sleeve 17 is made of rubber and elastic, one end of the communicating sleeve is sleeved at the free end of the air inlet pipe 18 of the air pump when the air pump 26 pumps air, the other end of the communicating sleeve is sleeved at the outer end of the air adjusting pipe 3, the inner diameter of the communicating sleeve 17 is 0.45-0.9 cm, the length of the communicating sleeve 17 is 10-20 cm, and the thickness of the wall of the communicating sleeve 17 is 0.5-1 mm. The constant temperature illumination incubator 19 is an incubator in which a plurality of adjustable micro-aerobic culture boxes 22 are placed, and provides specific temperature and illumination for the adjustable micro-aerobic culture boxes 22, and has a cuboid structure and functions similar to those of a well-known illumination incubator, wherein the cuboid has a length of 50-300 cm, a width of 50-100 cm and a height of 150-300 cm, and comprises a power plug 20 of the constant temperature illumination incubator, a power line 21 of the constant temperature illumination incubator, the adjustable micro-aerobic culture boxes 22, a light 23, a transparent support plate 24 and a cavity 25 of the constant temperature illumination incubator. The structure and function of the power plug 20 of the constant-temperature illumination incubator are the same as those of a well-known two-phase power plug, and electric energy can be led into the power cord 21 of the constant-temperature illumination incubator from the outside. The structure and the function of the power line 21 of the constant-temperature illumination incubator are the same as those of a well-known double-strand copper core power line, the cross section of the copper core is circular, the diameter of the circular copper core is 1-5 mm, and electric energy can be led into the constant-temperature illumination incubator 19 from a power plug 20 of the constant-temperature illumination incubator. The number of the light lamps 23 is 1-4, which are the same as the known fluorescent lamps in structure and function, are arranged at the top of the constant-temperature illumination incubator cavity 25 and can provide illumination for the adjustable micro-aerobic incubator 22. The transparent support plate 24 is made of glass or plastic with high transparency and is 0.3-0.8 cm thick, is arranged on a support in two side walls of the constant-temperature illumination incubator 19, can divide the constant-temperature illumination incubator cavity 25 into 2-5 parts, and can provide support for the adjustable micro-aerobic incubator 22. The constant-temperature illumination incubator cavity 25 is a space inside the constant-temperature illumination incubator 19, transparent support plates 24 are arranged in the constant-temperature illumination incubator cavity, the adjustable micro-aerobic culture boxes 22 are placed on the transparent support plates 24, and the transparent support plates 24 are made of glass or plastic with high transparency, so that the adjustable micro-aerobic culture boxes 22 placed on each transparent support plate 24 can be guaranteed to obtain certain illumination.

The foregoing shows and describes the general principles and features of the present invention, together with the advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are merely illustrative of the principles of the invention, but that various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined by the appended claims and their equivalents.

Claims

1. Be suitable for archaea culture apparatus of low temperature little oxygen environment growth, constitute its characterized in that by adjustable little oxygen culture box (22), constant temperature illumination incubator (19) and aspiration pump (26): the adjustable micro-aerobic culture box (22) is in a cuboid shape, the length of the cuboid is 15-20 cm, the width of the cuboid is 15-20 cm, and the height of the cuboid is 20-30 cm, the adjustable micro-aerobic culture box (22) comprises a water containing box (1), oxygen-controlled water (2), an air adjusting pipe (3), a scale oxygen-limiting cylinder (4), a foam buoyancy block (5), a culture dish (6), an air adjusting pipe leather plug (7), a water containing box wall (8), a water containing box cavity (9) and a micro-oxygen space (10), the oxygen-controlled water (2) is contained in the water containing box (1), the scale oxygen-limiting cylinder (4) is inserted into the oxygen-controlled water (2) in the water containing box (1) with an opening facing downwards, the micro-oxygen space (10) is formed at the bottom of the scale oxygen-limiting cylinder (4) as the oxygen-controlled water (2) is only partially filled in the scale oxygen-limiting cylinder (4), the culture dish (6) adhered to the foam buoyancy block (5) can be inoculated with the aid of the buoyancy block (5) after the bacteria are inoculated The oxygen control water is suspended on the upper surface of the oxygen control water (2) below the micro-oxygen space (10), one end of the air adjusting pipe (3) is communicated with the micro-oxygen space (10), the other end of the air adjusting pipe is blocked by an air adjusting pipe leather block (7), the air adjusting pipe leather block (7) is taken down to be communicated with the air extracting pump (26) during air adjusting, air in the micro-oxygen space (10) is extracted, and then the content of oxygen in the micro-oxygen space (10) is adjusted; the water containing box (1) is cuboid, the length of the cuboid is 15-20 cm, the width of the cuboid is 15-20 cm, and the height of the cuboid is 20-30 cm; the outer wall of the water containing box (1) is a water containing box wall (8), wherein the water containing box (1) is not provided with an upper bottom wall, the lower bottom and the water containing box walls (8) at the periphery are made of glass or plastics, and the thickness of the glass or plastics is 0.3-0.6 cm; the cavity surrounded by the water containing box walls (8) is a water containing box cavity (9); the oxygen control water (2) is tap water used in daily life; the air adjusting pipe (3) is made of glass or plastic and is U-shaped, the arm length of the U-shaped air adjusting pipe is 18-28 cm, the cross section of the air adjusting pipe (3) is circular, the outer diameter of the circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, the thickness of the circular ring is 0.1-0.2 cm, and the air adjusting pipe (3) is fixed on the wall of the scale oxygen limiting cylinder (4); the air adjusting pipe leather plug (7) is rubber and elastic, one end of the air adjusting pipe leather plug is opened, the other end of the air adjusting pipe leather plug is closed, the inner diameter of the air adjusting pipe leather plug is 0.45-0.9 cm, the length of the air adjusting pipe leather plug is 1-2 cm, the wall thickness of the air adjusting pipe leather plug is 0.5-1 mm, the air adjusting pipe leather plug is taken down from the air adjusting pipe (3) when the air pump (26) pumps air, and the air adjusting pipe leather plug is sleeved at the outer end of the air adjusting pipe (3) after air is pumped; the scale oxygen-limiting cylinder (4) is a container which limits the foam buoyancy block (5) and the culture dish (6) and provides a micro-oxygen environment for the cultured archaea in the culture dish (6), is cylindrical, has the diameter of 10-16 cm and the height of 18-28 cm, the upper bottom and the peripheral wall of the scale oxygen-limiting cylinder (4) are made of glass or plastic, the thickness of the scale oxygen-limiting cylinder is 1-2 mm, the scale oxygen-limiting cylinder (4) is not provided with the upper bottom wall, oxygen-controlling water (2), the foam buoyancy block (5) and the culture dish (6) enter the scale oxygen-limiting cylinder (4), volume scales are arranged on the peripheral wall of the scale oxygen-limiting cylinder (4) from top to bottom, and the volume scales are finished from 0 ml to 1000 ml; the foam buoyancy block (5) is a buoyancy part which is adhered to the lower part of the culture dish (6) and provides buoyancy for the culture dish (6) to enable the culture dish (6) to float on the liquid surface of the oxygen control water (2) in the scale oxygen limiting cylinder (4), is made of foam plastic and is in a round cake shape, and the diameter of the round cake is 5-12 cm, and the thickness of the round cake is 1-2 cm; the culture dish (6) is a part for culturing archaea, the part poured with the culture medium is a culture dish basal disc, and the part covering the culture dish basal disc is a culture dish cover; the culture medium used in the culture dish (6) is beef extract peptone culture medium or commercially available LB culture medium added with 1-5 ml of living environment leachate of a sample to be cultured; the micro-oxygen space (10) is a closed space formed by filling oxygen control water (2) into the scale oxygen limiting cylinder (4), the foam buoyancy block (5) and the culture dish (6) are suspended on the upper surface of the oxygen control water (2) in the micro-oxygen space (10), the size of the micro-oxygen space (10) changes along with the change of air suction amount, the volume scale on the wall of the scale oxygen limiting cylinder (4) can be read in real time, and the pressure in the micro-oxygen space (10) is controlled by the adding amount of the oxygen control water (2) and an operation mode.

2. An archaebacteria culture device suitable for growth in a low temperature micro-aerobic environment according to claim 1, wherein: the air pump (26) is an independent part for adjusting and controlling the size of the micro-oxygen space (10) by pumping gas in the micro-oxygen space (10) through the air adjusting pipe (3), and the structure and the function of the air pump (26) are the same as those of a known air pump and consist of an air pump power plug (11), an air pump power cord (12), an air pump handle (13), an air pump bracket (14), an air pump air outlet (15), an air pump body (16), a communicating sleeve (17) and an air pump air inlet pipe (18); the structure and the function of the power plug (11) of the air pump are the same as those of a well-known two-phase power plug, the structure and the function of the power cord (12) of the air pump are the same as those of a well-known two-strand copper core power cord, the cross section of the copper core is circular, the diameter of the circle is 1-5 mm, the handle (13) of the air pump is made of stainless steel or aluminum alloy, the cross section of the copper core is circular, the diameter of the circle is 1-3 cm, the length of the circle is 30-40 cm, the two ends of the circle are bent towards the same direction, the length of the bend is 5-10 cm, and the two ends of the bend are welded on the upper surface of the air pump body (16), so that a handle shape is formed; the air suction pump supports (14) are welded on the lower surface of the air suction pump body (16), 1 air suction pump support is arranged at each of the front end and the rear end of the air suction pump body (16), each air suction pump support (14) is made of stainless steel or aluminum alloy and is solid, the cross section of each air suction pump support is a trapezoid with a concave upper bottom surface, the length of the upper bottom edge of the trapezoid is 3-5 cm, the length of the lower bottom edge of the trapezoid is 10-15 cm, and the height of the trapezoid is 3-5 cm; the air outlet (15) of the air pump is an opening of the air pump body (16) which is discharged after the micro-oxygen space (10) is extracted by the air pump (26), and is positioned at one end of the air pump body (16), and an iron mesh is arranged at the opening; the air-extracting pump body (16) is cylindrical, the diameter of the cylinder is 10-15 cm, the length of the cylinder is 20-30 cm, and the structure and the function of the air-extracting pump body (16) are the same as those of a well-known air-extracting pump (26); the air intake pipe (18) of the air pump is a long pipe which is positioned at the other end of the air pump body (16) opposite to the air outlet (15) of the air pump and is made of stainless steel or aluminum alloy, the length of the air intake pipe is 10-20 cm, the cross section of the air intake pipe (18) of the air pump is circular, the outer diameter of the circular ring is 0.5-1 cm, the inner diameter of the circular ring is 0.3-0.6 cm, and the thickness of the circular ring is 0.1-0.2 cm; the communicating sleeve (17) is made of rubber and elastic, one end of the communicating sleeve is sleeved at the free end of the air inlet pipe (18) of the air suction pump when the air suction pump (26) sucks air, the other end of the communicating sleeve is sleeved at the outer end of the air adjusting pipe (3), the inner diameter of the communicating sleeve (17) is 0.45-0.9 cm, the length of the communicating sleeve (17) is 10-20 cm, and the thickness of the wall of the communicating sleeve (17) is 0.5-1 mm.

3. An archaeal culture device suitable for growth in a low temperature micro-aerobic environment according to claim 1, wherein: the constant-temperature illumination incubator (19) is cuboid, the length of the cuboid is 50-300 cm, the width of the cuboid is 50-100 cm, and the height of the cuboid is 150-300 cm, and comprises a power plug (20) of the constant-temperature illumination incubator, a power line (21) of the constant-temperature illumination incubator, an adjustable micro-aerobic incubator (22), a light (23), a transparent support plate (24) and a cavity (25) of the constant-temperature illumination incubator; the structure and the function of the power plug (20) of the constant-temperature illumination incubator are the same as those of a known two-phase power plug; the structure and the function of the power line (21) of the constant-temperature illumination incubator are the same as those of a known double-strand copper core power line, the cross section of the copper core is circular, and the diameter of the circle is 1-5 mm; the structure and the function of the illumination lamps (23) are the same as those of the well-known fluorescent lamps, 1 to 4, and the illumination lamps are arranged at the top of the constant-temperature illumination incubator cavity (25); the transparent support plate (24) is made of glass or plastic with higher transparency, has the thickness of 0.3-0.8 cm, and is arranged on the support in the two side walls of the constant-temperature illumination incubator (19); the constant-temperature illumination incubator cavity (25) is a space inside the constant-temperature illumination incubator (19), a transparent support plate (24) is arranged inside the constant-temperature illumination incubator cavity, and the adjustable micro-aerobic incubator (22) is placed on the transparent support plate (24).