CN110878252A

CN110878252A - Culture cylinder, culture system and culture method for microbial culture

Info

Publication number: CN110878252A
Application number: CN201911179553.7A
Authority: CN
Inventors: 肖英平; 吕文涛; 杨华; 徐娥; 王珍; 刘秀婷
Original assignee: Zhejiang Academy of Agricultural Sciences
Current assignee: Zhejiang Academy of Agricultural Sciences
Priority date: 2019-11-27
Filing date: 2019-11-27
Publication date: 2020-03-13
Anticipated expiration: 2039-11-27
Also published as: CN110878252B

Abstract

The invention relates to the technical field of microorganism culture, in particular to a culture cylinder, a culture system and a culture method for microorganism culture. The culture cylinder reduces the risk of death or variation or pollution in the growth and development propagation process of the microorganisms, ensures the good development speed of the microorganisms and saves the culture time.

Description

Culture cylinder, culture system and culture method for microbial culture

Technical Field

The invention relates to the technical field of microbial culture, in particular to a culture cylinder, a culture system and a culture method for microbial culture.

Background

In the case of microorganisms, which are a large group of organisms including bacteria, viruses, fungi, and some small protozoa, they are widely present in the biological world, and although individual microorganisms are small, they are closely related to human life. Is widely applied to the fields of medicine and health, industry and agriculture, environmental protection and the like.

Over the years, microbiologists have developed scientific tests from many aspects such as classification, physiology, heredity and the like, study the physiological and biochemical characteristics of microorganisms, and summarize a whole set of microorganism detection, screening and cultivation methods, so that microorganisms can be cultivated as required by people to serve the people.

The current microbial cultivation generally adopts the following steps:

firstly, collecting a microorganism specimen, then pretreating the collected microorganism specimen material to reduce the impurity content in the specimen material, then carrying out enrichment culture to obtain a large number of microorganisms, then selecting the microorganisms to obtain a primary selection strain, carrying out performance detection on the primary selection strain, judging whether each performance of the selected strain can meet the requirement of inoculation culture, and if the strain meets the requirement of inoculation culture, determining that the strain is qualified. After the qualified strains are obtained, the strains are preserved, so that the life activities of the microorganisms are in a semi-permanent dormant state, and when the strains need to be used, the corresponding strains are recovered.

Although the above-mentioned methods are widely used in the field of current microorganism application technology, in further research, the inventors found that the current methods still have disadvantages, such as the following:

although the preservation of microorganisms can be realized for a long time and the convenience of use is ensured, the preservation method still has the following defects: on one hand, a great deal of cost is consumed for strain preservation, and the cost of strain preservation occupies a great cost proportion in the current microbial application field; on the other hand, in the strain preservation process, preservation temperature, sealing quality and gas dryness all affect preservation quality, and when the conditions are changed, strains can die, change or be polluted; on the other hand, the recovery of the strains also needs to be carried out at a high cost, and in the recovery process of the strains, the risks of strain death and pollution also exist.

Therefore, there is a need to design a microbial cultivation system that can avoid the high cost of the strains due to the preservation and recovery processes and the risk of strain death and contamination.

Disclosure of Invention

The invention aims to: aiming at the defects that the cost is high due to a strain preservation and recovery process and the risk of strain death and pollution exists in the conventional microbial culture, the microbial culture system capable of avoiding the high cost of the strain due to the strain preservation and recovery process and the risk of strain death and pollution is provided.

In order to achieve the above purpose, the invention provides the following technical scheme:

the utility model provides a cultivate section of thick bamboo for microbial cultivation, including the lower extreme seal the open barrel in upper end and with the open tip matched with lid in barrel upper end the barrel inner space divides culture medium chamber and gas chamber from bottom to top, the culture medium chamber is used for holding the culture medium that suitable microorganism grows, the gas chamber is used for holding the suitable gas that suitable microorganism grows still be provided with gaseous replacement device in the barrel, gaseous replacement device is used for making the gas in the gas chamber is replaced for new suitable gas.

In the scheme of the present application, the culture medium suitable for culturing the microorganism is a conventional culture medium at present, and may be a liquid culture medium, a solid culture medium or a culture medium in a colloid state, and for different types of microorganisms, the formula and the form of the culture medium are different, and for different types of microorganisms, the formula of the gas suitable for propagation and growth of the microorganism may also be different, and the formula, the form of the culture medium, and the suitable gas of the microorganism are conventional techniques in the art that are known by those skilled in the art, and thus suitable culture medium and gas suitable for the microorganism can be selected or prepared for different types of microorganisms according to the general techniques in the art, and thus, details are not repeated in this content.

In the current culture of microorganisms, periodic gas components are critical factors influencing the propagation, growth and development of microorganisms, when the ambient gas components are greatly changed, the propagation, growth and development of microorganisms are influenced, and even the microorganisms die or change seriously, or cause pollution and other problems, which are also the key aims of the current strain preservation, in the practical work, the inventor finds that during the growth and development of the microorganisms, no matter aerobic microorganisms or anaerobic microorganisms, most of the microorganisms influence the ambient air, the growth and development of some microorganisms absorb partial components in the ambient air, some microorganisms release some gases, some microorganisms absorb certain gases and also discharge some gases, so the ambient gas of the microorganisms also changes along with the growth and development of the microorganisms over time, this results in the risk of death, mutation and contamination of the microorganisms during their growth and development due to the change in gas composition. Based on the above, in the scheme of this application, the microorganism is cultivateed in the culture medium in the barrel, and the gas chamber holds suitable gas, and as time goes on, when the gas in the gas chamber no longer is suitable for the microorganism to grow the reproduction development, through the gas replacement device in the barrel, replace the gas in the gas chamber with new suitable gas, so, reduce microorganism growth reproduction process death or variation or appear contaminated risk, moreover, also guaranteed the good developmental rate of microorganism, practiced thrift the culture time.

The application also discloses a gaseous replacement device for above-mentioned culture section of thick bamboo, gaseous replacement device is including replacing subassembly and gaseous exhaust subassembly, gaseous exhaust subassembly is used for the intercommunication the exterior space of gas chamber and barrel, the replacement subassembly is used for promoting gaseous intracavity makes gaseous intracavity gaseous by gaseous exhaust subassembly discharges outside the barrel.

As the preferred technical scheme, the gas discharge assembly comprises a communicating pipe, one end of the communicating pipe is communicated with the gas cavity, and the other end of the communicating pipe penetrates through the cylinder body and is communicated with the external space of the cylinder body.

As a preferable technical scheme, a stop valve is further arranged on the communicating pipe. And the communication and blocking states of the communication pipe are controlled by opening and closing the stop valve.

As another preferable technical solution, a one-way conduction device is further disposed on the communication pipe, so that the communication pipe is in one-way conduction in a direction from the inside of the barrel body to the outside of the barrel body.

As a preferred technical scheme, the one-way conduction device is arranged at one end, close to the gas cavity, of the communication pipe.

Preferably, the one-way conduction device is a one-way valve.

As a preferable technical scheme, the communicating pipe is vertically arranged along the inner wall of the gas cavity, the lower end of the communicating pipe is in clearance fit with the upper edge of the culture medium cavity, and the upper end of the communicating pipe penetrates through the upper end part of the cylinder body to form the exhaust nozzle.

As another preferred technical scheme, the communicating pipe comprises a penetrating section and a vertical section, the vertical section is vertically arranged along the outer wall of the cylinder, one end of the penetrating section is positioned in the cylinder and is in clearance fit with the upper edge of the culture medium cavity, and the other end of the penetrating section penetrates out of the cylinder and is connected with the lower end of the vertical section.

As a further preferable technical solution, the piercing section is inclined downward in a direction from inside the barrel to outside the barrel. The penetration section is inclined downwards, and as mentioned above, when the exhaust gas is condensed into liquid drop particles in the communicating pipe, the particles can be prevented from falling into the culture substrate cavity, and the good culture quality of the microorganism is ensured.

As a further preferable technical scheme, a downward bent U-shaped bent pipe is connected between the piercing section and the vertical section.

As a further preferable technical scheme, a drain valve is further arranged at the bottom of the U-shaped elbow. When needed, the drain valve can be opened to drain liquid and/or solid particles in the elbow.

As a preferred technical scheme, the replacement component comprises a push plate and a push rod arranged on the upper side of the push plate, the push rod is vertically arranged and penetrates through the cover body, the push rod is in sliding sealing fit with the cover body, and the push plate is in sliding sealing fit with the side wall of the gas cavity.

As the preferred technical scheme, work as communicating pipe is located when the barrel, communicating pipe corresponds be provided with the breach on the push pedal, the breach of push pedal with be sliding seal cooperation between communicating pipe.

According to the preferable technical scheme, a through hole penetrating through the push rod from top to bottom is formed in the push rod, the through hole penetrates through the push plate, and a blocking cover used for blocking the through hole is detachably arranged at the upper end of the push rod.

As a preferred technical scheme, the lower side surface of the push plate is in a conical shape with the middle part protruding upwards, and the through hole of the push rod is positioned at the top of the conical surface on the lower side of the push plate. The lower side surface of the push plate is arranged to be in a conical shape with the middle part protruding upwards, when a liquid culture medium enters the through hole and flows downwards to the lower side of the push plate, the liquid culture medium can flow downwards in a radial mode along the conical surface, and then the uniformity of liquid dispersion of the added culture medium is improved.

As a further preferred technical scheme, a plurality of guide grooves along the direction of a generatrix are arranged on the lower side of the push plate, and the guide grooves are uniformly distributed on the circumference. Through the arrangement of the diversion trenches, the dispersion uniformity of the culture medium liquid in the circumferential direction is further improved.

As a further preferable technical scheme, a spray head connected with the through hole is further arranged on the lower side of the push plate corresponding to the through hole, and the spray head is used for spraying the liquid culture medium into a mist shape. When the culture substrate is liquid, the supplemented culture substrate liquid is sprayed into a mist shape by arranging the spray head, and the mist liquid particles are gradually settled in the culture substrate cavity, so that the uniformity of dispersion is ensured, and the culture quality of microorganisms is further ensured.

As a preferred technical scheme, the side wall of the cylinder is also provided with an observation window, the observation window is arranged on the side wall of the cylinder corresponding to the gas cavity, the edge of the observation window is in sealing fit with the edge of the cylinder, and the observation window is made of transparent materials. The upper opening of the barrel is provided with the observation window, so that the microorganism condition and the condition of each component in the barrel can be observed conveniently and visually, and the operation of operators is facilitated.

Preferably, the cylinder is further provided with a barometer for monitoring the air pressure in the air cavity. Through the setting of barometer, the control to the gas cavity internal gas pressure is convenient, simultaneously, also makes things convenient for the control to the volume of supplementing gas.

The application also discloses a microbial cultivation system, including foretell microbial cultivation section of thick bamboo, still include the control by temperature change cabin, the control by temperature change under-deck has one and can keep the stable temperature control chamber of cavity internal temperature, the whole or lower part of a cultivation section of thick bamboo is so that in the temperature control chamber. In the microbial culture, the temperature of the culture medium and the environment around the microbes is also one of important factors, whether the temperature is proper or not is determined to determine the growth and development quality of the microbes, in the actual microbial culture, some microbes can absorb heat to cause the temperature of a culture substrate part to be reduced, some microbes can emit heat to cause the temperature of the culture substrate part to be increased, and the temperature change often inhibits the further growth and development of the microbes, and in severe cases, the microbes can even have variation and death.

Therefore, in the scheme of this application, through setting up the control by temperature change cabin, the temperature in the temperature control intracavity is controllable adjustable to can keep the temperature stable in the cavity, so, make the temperature in the culture section of thick bamboo, especially the temperature in culture medium chamber all be in the temperature that is fit for the microorganism growth and development reproduction, and then ensure the good cultivation quality of microorganism.

As a preferred technical scheme, the temperature control cabin is a closed cavity, a heat transfer object is filled in the closed cavity, the temperature control cabin is also provided with a temperature adjusting device for heating and/or refrigerating the heat transfer object, a concave cavity which is concave towards the cabin body is further arranged on an upper side plate of the temperature control cabin, the culture cylinder is arranged in the concave cavity, and the side wall of the concave cavity is matched with the outer wall of the culture cylinder. In this kind of mode, through setting up attemperator to suitable temperature, through setting up the heat conduction thing in closed intracavity, the heat conduction thing can be water, oil etc. can realize the material to sunken intracavity uniform heating, also can be the air, so realize controlling the temperature in the sunken intracavity, and then realize the control to the temperature in the microorganism culture section of thick bamboo.

As a preferable technical scheme, a flexible sealing gasket is further arranged on the upper edge of the concave cavity and used for sealing a gap between the outer wall of the culture cylinder and the concave cavity. The setting of sealed pad is on the one hand to ensure good heat transfer between sunken chamber and the barrel, and on the other hand has also improved the stability of barrel.

Preferably, the sealing gasket is a rubber sealing gasket.

As a preferable technical scheme, a plurality of concave cavities are arranged on the temperature control cabin, and each concave cavity is internally provided with one culture cylinder. The culture efficiency can be greatly improved, the number of microorganisms in a single cylinder can be greatly reduced when the same number of microorganisms are cultured, the culture quality is improved, the risk of cross infection among the microorganisms is greatly reduced, and the mutation control of the microorganisms is also utilized.

Preferably, the adjacent concave cavities are separated by a distance, so that a channel for people to pass through is formed between the culture cylinders after the culture cylinders are arranged in the concave cavities. The staff is convenient to monitor and operate each culture cylinder.

As a preferred technical scheme, the microorganism culture system also comprises a box body, the temperature control cabin and the culture cylinder are arranged in the box body, and the temperature control cabin is detachably connected with the box body. Through setting up the box, improve the wholeness of culture system, when reducing the pollution that the culture section of thick bamboo receives, still convenient transportation.

Preferably, the container body is a container.

As a preferable technical scheme, a gas chamber is further arranged in the box body and used for storing gas suitable for the cultured microorganisms, and a gas filling pipeline matched with the through hole of the push rod of the culture cylinder is further arranged on the gas chamber. The gas chamber has a closed cavity within which new unused gas is stored. The air charging pipeline is also provided with an air charging nozzle matched with the pushing through hole, and the air charging nozzle is in separable sealing fit with the push rod. After the air in the gas cavity in the culture cylinder is exhausted, the gas cavity is filled with new suitable gas suitable for the growth and development of microorganisms through the matching of the gas-filling pipeline and the through hole of the push rod.

Preferably, the gas chamber is disposed at a top of the tank.

According to a preferable technical scheme, the gas chamber comprises a lower side plate, the edge of the lower side plate is in sliding sealing fit with the side wall of the box body, a gas storage space of the gas chamber is formed between the lower side plate and a top plate of the box body, the gas adding pipe is arranged on the lower side plate and communicated with the gas storage space of the gas chamber, and a driving device for driving the lower side plate to move vertically is further arranged in the box body. The driving device is a driving motor, the lower side plate is driven to move upwards through the driving motor, and gas in the gas chamber is pressed into the gas cavity of the culture cylinder, so that the suitable gas is supplemented.

Preferably, an exhaust gas chamber is further provided in the case, and the exhaust gas chamber is communicated with the communication pipe of each of the culture cylinders. In response to certain microbial cultures, suitable gases in the gas chamber may cause serious pollution or toxic or harmful gases, and in response to such toxic or harmful gases, in the scheme of the application, the cylinder waste gas chamber is collected and then subjected to centralized harmless treatment.

The application also discloses a microbial culture method adopting the microbial culture system, after microbial strains are obtained, the strains are cultured in the microbial culture system, then the microbial culture system with the microbial strains is transported to a use site, and then the microbes in the culture system are taken out for on-site use.

Compared with the traditional mode, the microbial culture method directly avoids the steps of preserving microbial strains and recovering the microbial strains in the traditional mode because the microbial culture system is adopted in the transportation link, so that the preservation and recovery cost is firstly avoided, more importantly, the risks of death and variation of the microbes caused in the traditional preservation and recovery process are avoided, and by adopting the mode of the application, the microbes directly enter the culture system to culture after being cultured, the microbial variation risk is reduced, the purity of the microbes is improved, and the microbes have excellent activity and are developed and propagated to a certain quantity when being transported to a use site for use, the time period of field use is greatly shortened, and the use cost is greatly reduced, the microorganism culture method is particularly suitable for the situation that the microorganism needs to be used immediately or in a short time at the use site.

As a preferable technical scheme, after the microbial strains are obtained, before the strains are cultured in a microbial culture system, the gas replacement of the microbial culture system and the time node experiment of culture medium supplement are firstly carried out,

the microorganism culture system comprises a gas replacement and culture substrate supplement time node experiment: setting the temperature control cabin to be suitable for the growth, development and propagation of microorganisms, placing a culture substrate and strains into a culture substrate cavity of the culture cylinder according to the filling amount of the culture substrate and the placing amount of the microorganism strains in the culture cylinder in the actual transportation process, then filling suitable gas into a gas cavity of the culture cylinder, starting timing, recording a time reading h0, selecting a microorganism mutation rate threshold value and a death rate threshold value according to the requirements of actual microorganism strains and the use field, and then carrying out the following steps:

step 1: monitoring the microorganisms in the culture medium, recording a time reading h1 when any one of the mutation rate and the death rate of the microorganisms in the culture cylinder reaches 90-95% of a threshold value,

step 2: detecting the components and the proportion of each component of the culture medium at h1, comparing the proportion of the components with the proportion of the original components of the culture medium, calculating the amount d1 of the components to be supplemented,

and step 3: the amount of the supplementary ingredient d1 is added into the culture substrate chamber, then the gas in the gas chamber is replaced by new suitable gas,

and 4, step 4: continuing to monitor the variation rate and the mortality rate of the microorganisms, recording the time reading h2 when any one of the variation rate and the mortality rate of the microorganisms in the culture cylinder reaches 90-95% of the threshold value,

and 5: detecting the components and the proportion of each component of the culture medium at h2, comparing the proportion of the components with the proportion of the original components of the culture medium, calculating the amount d2 of the components to be supplemented,

step 6: the amount of the supplementary ingredient d2 is added into the culture substrate chamber, then the gas in the gas chamber is replaced by new suitable gas,

repeating the steps 3-6 to obtain data h3-hn and d3-dn, n is an integer greater than three,

when the strain is cultured in a microorganism culture system, according to the time h1-hn, when the time reaches a certain hx, the corresponding component amount dx is supplemented into a culture medium cavity of a culture cylinder, and meanwhile, the gas in a gas cavity of the culture cylinder is replaced by new suitable gas, wherein x is more than or equal to 1 and less than or equal to n, and x is an integer.

In the above-mentioned mode of the present application, the microorganism monitoring is performed by using the conventional microorganism monitoring mode, the specific monitoring process can be obtained by those skilled in the art without any doubt by using conventional experimental equipment and conventional technical means, and the experiments are not repeated in the present application, and in the actual culture process, the inventors found that, because the microorganism environmental gas not only contains the gas discharged during the growth process of the microorganism but also contains the gas volatilized by the culture substrate, and the absorption and use of the components of the culture substrate by the microorganism are generally less, and the change of the gas components within the same time is larger, and the change of the components of the microorganism is more significant compared with the change of the components of the culture substrate, in the present scheme, the microorganism variation threshold and the death threshold are selected according to the corresponding standards and specifications of different microorganism varieties, and through each time node, in the culture system, and gas replacement and corresponding culture medium supplement are carried out according to time nodes, so that the whole culture process is ensured, and the microorganisms are all in a proper environment, and the culture quality of the microorganisms is ensured.

As a preferable technical scheme, before the transportation of the microorganism culture system, the transportation time is estimated, the required appropriate gas volume and the required supplementary component amount before the transportation to the use site are calculated according to the transportation time and the number of culture cylinders in the microorganism culture system, and the required appropriate gas volume and the required supplementary component amount are arranged in a box body of the microorganism culture system.

Preferably, the amounts of the medium-supplemented components required by the culture system are grouped according to the actual data h, the amounts of the medium-supplemented components required by the entire microorganism culture system are grouped according to different time points, and the amounts of the medium-supplemented components required by the entire microorganism culture system are further equally divided according to the number of culture tubes and are divided into supplemented component groups corresponding to the culture tubes.

Preferably, the culture system is transported to a use site, and then the microorganism in the culture cylinder is mixed with the culture substrate and mixed with the product to be used.

Compared with the prior art, the invention has the beneficial effects that:

1. according to the culture cylinder, microorganisms are cultured in a culture medium in the cylinder body, the gas cavity contains suitable gas, and when the gas in the gas cavity is no longer suitable for the growth, the propagation and the development of the microorganisms, the gas in the gas cavity is replaced by new suitable gas through the gas replacement device in the cylinder body, so that the risk of death or variation or pollution in the growth, the development and the propagation processes of the microorganisms is reduced, the good development speed of the microorganisms is ensured, and the culture time is saved;

2. the unidirectional conducting device is arranged, so that the unidirectional conducting device only performs exhaust, and thus, adverse effects on microorganisms caused by that sediment in exhaust is brought into the cylinder again by newly introducing proper gas are avoided, and the culture quality of the microorganisms is further improved;

3. the U-shaped bent pipe is arranged, so that the gathering of condensed particles in the exhaust gas is facilitated, and the particles are brought back to the culture medium cavity by turbulent flow gas formed by the communicating pipe in the exhaust process;

4. the through holes are formed in the push rod in such a way that after the gas in the gas cavity is exhausted, new proper gas can be supplemented into the gas cavity through the through holes of the push rod, and in such a way, as mentioned above, the exhausted gas and the inlet gas respectively adopt different channels, so that the liquid condensed in the exhausted gas and/or fixed particles can be prevented from entering the culture substrate cavity as much as possible, and the culture quality of microorganisms is further ensured;

on the other hand, the component quality of the culture substrate is also an important factor influencing the culture quality of microorganisms, in the process of growth, development and propagation of the microorganisms, on one hand, the absorption of the microorganisms is realized, so that part of components in a culture medium liquid is consumed, and on the other hand, because the components of the culture substrate are isolated, especially a liquid culture medium, and after standing for a long time, part of components are settled in the lower part of liquid, the change of the components of the culture medium liquid is caused, the growth and development of the microorganisms are slow, and even the microorganisms are dormant or mutated in severe cases;

5. according to the microorganism culture system, in microorganism culture, the temperature of a culture medium and the environment around the microorganism is also one of important factors, whether the temperature is proper or not is determined, the growth and development quality of the microorganism is determined, in actual microorganism culture, some microorganisms can absorb heat to cause the temperature of a culture matrix part to be reduced, some microorganisms can emit heat to cause the temperature of the culture matrix part to be increased, the temperature change can often inhibit the further growth and development of the microorganism, and even mutation and death can occur in serious cases; therefore, in the scheme of the application, the temperature in the temperature control cavity is controllable and adjustable by arranging the temperature control cabin, and the temperature in the cavity can be kept stable, so that the temperature in the culture cylinder, particularly the temperature in the culture medium cavity is kept at the temperature suitable for the growth, development and propagation of microorganisms, and the good culture quality of the microorganisms is further ensured;

6. compared with the traditional mode, the microbial culture method directly avoids the steps of preserving microbial strains and recovering the microbial strains in the traditional mode because the microbial culture system is adopted in the transportation link, so that the preservation and recovery cost is firstly avoided, more importantly, the risks of death and variation of the microbes caused in the traditional preservation and recovery process are avoided, and by adopting the mode of the application, the microbes directly enter the culture system to culture after being cultured, the microbial variation risk is reduced, the purity of the microbes is improved, and the microbes have excellent activity and are developed and propagated to a certain quantity when being transported to a use site for use, the time period of field use is greatly shortened, and the use cost is greatly reduced, the microorganism culture method is particularly suitable for the situation that the microorganism needs to be used immediately or in a short time on the use site;

7. the method is characterized in that a microorganism mutation rate threshold value and a microorganism death rate threshold value are selected according to corresponding standards and specifications of different microorganism varieties, and gas replacement and corresponding culture medium supplement are carried out according to time nodes in a culture system through each time node, so that the whole culture process is ensured, microorganisms are in a proper environment, and the culture quality of the microorganisms is ensured.

Description of the drawings:

FIG. 1 is a schematic structural view of a cartridge in an embodiment;

FIG. 2 is a schematic structural view of the temperature control chamber matched with the barrel and the box body in the specific embodiment,

the following are marked in the figure: 1-cylinder, 2-cover, 3-culture medium cavity, 4-gas cavity, 5-communicating pipe, 6-one-way conducting device, 7-U-shaped bent pipe, 8-push plate, 9-push rod, 10-through hole, 11-blocking cover, 12-diversion trench, 13-spray head, 14-observation window, 15-temperature control cabin, 16-temperature control cavity, 17-sealing gasket, 18-box, 19-gas chamber, 20-waste gas chamber.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments.

Thus, the following detailed description of the embodiments of the invention is not intended to limit the scope of the invention as claimed, but is merely representative of some embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that the embodiments of the present invention and the features and technical solutions thereof may be combined with each other without conflict.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

In the description of the present invention, it should be noted that the terms "upper", "lower", and the like refer to orientations or positional relationships based on those shown in the drawings, or orientations or positional relationships that are conventionally arranged when the products of the present invention are used, or orientations or positional relationships that are conventionally understood by those skilled in the art, and such terms are used for convenience of description and simplification of the description, and do not refer to or imply that the devices or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like are used merely to distinguish one description from another, and are not to be construed as indicating or implying relative importance.

Example 1, as shown in figures 1 and 2:

the utility model provides a be used for microorganism cultivation section of thick bamboo, including the lower extreme seal the open barrel 1 in upper end and with 1 upper end of the barrel opens tip matched with lid 2 1 inner space from bottom to top divide into culture medium chamber 3 and gas chamber 4, culture medium chamber 3 is used for holding the culture medium that suitable microorganism grows, gas chamber 4 is used for holding the suitable gas that suitable microorganism grows still be provided with gaseous replacement device in the barrel 1, gaseous replacement device is used for making gaseous replacement in the gas chamber 4 is new suitable gas.

In the current culture of microorganisms, periodic gas components are critical factors influencing the propagation, growth and development of microorganisms, when the ambient gas components are greatly changed, the propagation, growth and development of microorganisms are influenced, and even the microorganisms die or change seriously, or cause pollution and other problems, which are also the key aims of the current strain preservation, in the practical work, the inventor finds that during the growth and development of the microorganisms, no matter aerobic microorganisms or anaerobic microorganisms, most of the microorganisms influence the ambient air, the growth and development of some microorganisms absorb partial components in the ambient air, some microorganisms release some gases, some microorganisms absorb certain gases and also discharge some gases, so the ambient gas of the microorganisms also changes along with the growth and development of the microorganisms over time, this results in the risk of death, mutation and contamination of the microorganisms during their growth and development due to the change in gas composition. Based on the above, in the scheme of this application, the microorganism is cultivated in the culture medium in barrel 1, and gas chamber 4 holds suitable gas, and as time goes on, when the gas in gas chamber 4 no longer suits the growth, reproduction and development of microorganism, through the gas replacement device in barrel 1, replace the gas in gas chamber 4 with new suitable gas, so, reduce the risk that microorganism growth, development and propagation process dies or mutates or appears contaminated, and, also, guaranteed the good development speed of microorganism, practiced thrift culture time.

Example 2, as shown in figures 1 and 2:

a gas replacement device for the culture cylinder of the embodiment comprises a replacement component and a gas discharge component, wherein the gas discharge component is used for communicating the gas cavity 4 with the external space of the cylinder body 1, and the replacement component is used for pushing the gas in the gas cavity 4, so that the gas in the gas cavity 4 is discharged out of the cylinder body 1 through the gas discharge component.

As a preferred embodiment, the gas discharge assembly includes a communication pipe 5, one end of the communication pipe 5 is communicated with the gas chamber 4, and the other end of the communication pipe 5 passes through the cylinder 1 to be communicated with the external space of the cylinder 1.

In a preferred embodiment, a shutoff valve is further provided in the communication pipe 5. The communication and blocking states of the communication pipe 5 are controlled by the opening and closing of the shutoff valve. Through setting up the stop valve, the stop valve is normally closed state, when needing the gaseous in the replacement gas chamber 4, opens the stop valve, promotes gaseous in the gas chamber 4 through replacing the subassembly, makes gaseous 4 interior gas by communicating pipe 5 discharge, then fills new suitable gas in 5 towards the gas chamber 4 through communicating pipe, so realizes gaseous replacement.

As another preferred embodiment, the communication pipe 5 is further provided with a one-way conduction device 6 for making the communication pipe 5 conduct in one direction from the inside of the cylindrical body 1 to the outside of the cylindrical body 1. So that air outside the cylinder 1 cannot enter the gas chamber 4 through the blocking assembly. In practical use, the inventor finds that, because the gas in the gas chamber 4 is usually relatively humid, after the communicating pipe 5 exhausts for many times, there is a high possibility that liquid droplet particles are gathered in the communicating pipe 5, and even after a longer time, the particles are dried into solid or dust particles, if the communicating pipe 5 is adopted to blow new suitable gas into the gas chamber 4, the particles can be sent back into the gas chamber 4 and fall onto the culture medium, and abnormal substances with high concentration are formed on the particles and the culture medium around the particles, which can cause local microorganism mutation and death very easily, and further affect the culture effect of the whole microorganism, therefore, in the scheme of the application, the one-way conduction device 6 is arranged, so that the one-way conduction device 6 only exhausts gas, and thus, the adverse effect on the microorganism caused by the sediment during exhausting being brought into the cylinder 1 by the new suitable gas is avoided, thereby providing the culture quality of the microorganism.

In a preferred embodiment, the one-way conducting device 6 is disposed at one end of the communicating pipe 5 close to the gas chamber 4. In this way, further ensure that the pollutant of communicating pipe 5 does not enter barrel 1, simultaneously, still ensure the gaseous discharge as much as possible in the gas chamber 4, avoid the residual gas in the pipeline to return to in the gas chamber 4.

In a preferred embodiment, the one-way conduction device 6 is a one-way valve.

As a preferred embodiment, the communicating tube 5 is vertically arranged along the inner wall of the gas chamber 4, the lower end of the communicating tube 5 is in clearance fit with the upper edge of the culture medium chamber 3, and the upper end of the communicating tube 5 penetrates through the upper end part of the cylinder 1 to form an exhaust nozzle. The communicating pipe 5 is vertically arranged on the side wall of the gas cavity 4, the exhaust nozzle is positioned at the upper end part of the cylinder body 1, so that the exhaust nozzle has larger operation space, and the phenomenon that a plurality of cylinder bodies 1 are separated by more distance for reserving the operation space when the exhaust nozzle is lower is avoided; moreover, 5 ways of communicating pipe are arranged on the inner wall of the gas cavity 4, so that the temperature of the communicating pipe 5 is consistent with the temperature in the gas cavity 4, the possibility that the gas is condensed into liquid drops when entering the communicating pipe 5 is reduced, and the pollution risk of the discharged gas to the communicating pipe 5 is further reduced.

As another preferred embodiment, the communicating tube 5 includes a penetrating section and a vertical section, the vertical section is vertically arranged along the outer wall of the cylinder 1, one end of the penetrating section is located in the cylinder 1 and is in clearance fit with the upper edge of the culture medium chamber 3, and the other end of the penetrating section penetrates out of the cylinder 1 and is connected with the lower end of the vertical section. In this scheme, set up the vertical section of communicating pipe 5 outside barrel 1, can effectively avoid the occupation to 1 inner space of barrel, also can ensure the level and smooth of 1 inside of barrel, reduce the gap that exists in the barrel 1 etc. reduce culture medium and assemble in gap department and lead to the problem that high concentration culture medium appears in this local department, and then avoided these high concentration culture medium to fall into below culture medium and the adverse effect that leads to.

As a further preferred embodiment, the piercing section is inclined downwards in the direction from inside the barrel 1 to outside the barrel 1. The piercing section is inclined downwards, as mentioned above, when the exhaust gas is condensed into droplet particles in the communicating tube 5, these particles can be prevented from falling into the culture substrate chamber 3, and the good culture quality of the microorganism is ensured.

As a further preferred embodiment, a downwardly bent U-bend 8 is connected between the piercing section and the vertical section. Through setting up U type return bend 8, do benefit to the gathering of the particulate matter that condenses in the exhaust gas more, reduce the exhaust in-process, the vortex gas that communicating pipe 5 formed takes the particulate matter back to culture medium chamber 3 in.

As a further preferred embodiment, a blowdown valve is further arranged at the bottom of the U-shaped elbow 8. When needed, the drain valve can be opened to drain the liquid and/or solid particles in the elbow 8.

As a preferred embodiment, the replacement assembly includes a push plate and a push rod 9 disposed on the upper side of the push plate, the push rod 9 is disposed vertically and passes through the cover 2, the push rod 9 and the cover 2 are in sliding sealing fit, and the push plate and the side wall of the gas cavity 4 are in sliding sealing fit. In this scheme, push rod 9 can be that manpower drive also can be set up to motor drive, through setting up push pedal and push rod 9, at initial position, the push pedal is located 4 tops of gas chamber, when needs carry out gas outgoing, promotes the push pedal through push rod 9, makes the push pedal move down, and the gas propelling movement in 4 gas chambers is sent into the gas outgoing subassembly, and then realizes gaseous discharge, and under this kind of mode, gas chamber 4 can be mended from communicating pipe 5 to new gas.

As a preferred embodiment, when the communicating pipe 5 is located in the cylinder 1, a notch is provided on the push plate corresponding to the communicating pipe 5, and the notch of the push plate is in sliding sealing fit with the communicating pipe 5. Set up in communicating pipe 5 complex breach on the push pedal, still make the part that communicating pipe 5 is located barrel 1 do guide structure, avoid the circumferencial direction's of push pedal gyration, reduce the wearing and tearing that the push pedal edge received, ensure the sealing reliability of push pedal and gas chamber 4 lateral walls.

As a preferred embodiment, a through hole 10 penetrating through the push rod 9 from top to bottom is arranged in the push rod 9, the through hole 10 penetrates through the push plate, and a blocking cover 11 for blocking the through hole 10 is detachably arranged at the upper end of the push rod 9. The through hole 10 is arranged on the push rod 9 in such a way that after the gas in the gas cavity 4 is exhausted, new suitable gas can be supplemented into the gas cavity 4 through the through hole 10 of the push rod 9, and in such a way, as mentioned above, the exhausted gas and the inlet gas respectively adopt different channels, so that the liquid and/or fixed particles condensed in the exhausted gas can be prevented from entering the culture substrate cavity 3 as much as possible, and the culture quality of microorganisms is further ensured;

on the other hand, the component quality of the culture substrate is also an important factor influencing the culture quality of microorganisms, in the process of growth, development and propagation of microorganisms, on one hand, the absorption of microorganisms is carried out, so that part of components in the culture medium liquid is consumed, on the other hand, because the components of the culture substrate are separated, especially the liquid culture medium, and part of components are settled in the lower part of liquid after being stood for a long time, the change of the components of the culture medium liquid is caused, the growth and development of the microorganisms are slow, and even the microorganisms are dormant or mutated in severe cases, in the scheme of the application, corresponding components can be supplemented into the culture substrate cavity 3 through the through hole 10 of the push rod 9, so that the components of the culture substrate are ensured to be in the component state suitable for the growth and development of the microorganisms, and the culture quality of the microorganisms is further reliably ensured.

In a preferred embodiment, the lower side surface of the push plate is in a shape of a cone with an upward convex middle part, and the through hole 10 of the push rod 9 is located at the top of the conical surface on the lower side of the push plate. The lower side surface of the push plate is arranged to be in a conical shape with the middle part protruding upwards, when a liquid culture medium enters the through hole 10 and flows downwards to the lower side of the push plate, the liquid culture medium can flow downwards along the conical surface in a radial shape, and then the uniformity of liquid dispersion of the added culture medium is improved.

As a further preferred embodiment, a plurality of flow guide grooves 12 are arranged on the lower side of the push plate along the generatrix direction, and the flow guide grooves 12 are uniformly distributed on the circumference. The distribution uniformity of the culture medium liquid in the circumferential direction is further improved by the arrangement of the diversion trenches 12.

In a further preferred embodiment, a spray head 13 connected to the through hole 10 is further disposed at the lower side of the push plate corresponding to the through hole 10, and the spray head 13 is used for spraying the liquid culture medium into a mist. When the culture substrate is liquid, the supplemented culture substrate liquid is sprayed into a mist shape by arranging the spray head 13, and the mist liquid particles are gradually settled in the culture substrate cavity 3, so that the uniformity of dispersion is ensured, and the culture quality of microorganisms is further ensured.

In a preferred embodiment, an observation window 14 is further disposed on a side wall of the cylinder 1, the observation window 14 is disposed on a side wall of the cylinder 1 corresponding to the gas chamber 4, an edge of the observation window 14 is in sealing fit with an edge of the cylinder 1, and the observation window 14 is made of a transparent material. The upper opening of the cylinder body 1 is provided with the observation window, so that the microorganism condition and the condition of each component in the cylinder body 1 can be conveniently and visually observed, and the operation of an operator is facilitated.

In a preferred embodiment, an air pressure gauge for monitoring the air pressure in the air chamber 4 is further provided on the cylinder 1. Through the setting of barometer, the control to the air pressure in gas chamber 4 is convenient, simultaneously, also convenient the control to the volume of supplementing gas.

Example 3, as shown in figures 1 and 2:

a microbial cultivation system comprises the microbial cultivation cylinder and a temperature control cabin 15, wherein a temperature control cavity 16 capable of keeping the temperature in the cavity stable is arranged in the temperature control cabin 15, and all or part of the cultivation cylinder is arranged in the temperature control cavity 16. In the microbial culture, the temperature of the culture medium and the environment around the microbes is also one of important factors, whether the temperature is proper or not is determined to determine the growth and development quality of the microbes, in the actual microbial culture, some microbes can absorb heat to cause the temperature of a culture substrate part to be reduced, some microbes can emit heat to cause the temperature of the culture substrate part to be increased, and the temperature change often inhibits the further growth and development of the microbes, and in severe cases, the microbes can even have variation and death.

Therefore, in the scheme of the application, the temperature in the temperature control cavity 16 can be controlled and adjusted by arranging the temperature control cabin 15, and the temperature in the cavity can be kept stable, so that the temperature in the culture cylinder, particularly the temperature in the culture medium cavity 3 is kept at the temperature suitable for the growth, development and propagation of microorganisms, and the good culture quality of the microorganisms is further ensured.

As a preferred embodiment, the temperature control cabin 15 is a closed cavity, a heat transfer object is filled in the closed cavity, the temperature control cabin 15 is further provided with a temperature adjusting device for heating and/or cooling the heat transfer object, a concave cavity which is concave into the cabin body is further arranged on an upper side plate of the temperature control cabin 15, the culture cylinder is arranged in the concave cavity, and a side wall of the concave cavity is matched with an outer wall of the culture cylinder. The sunken cavity is temperature control chamber 16, and in this kind of mode, through setting up attemperator to suitable temperature, through setting up the heat-transfer thing in closed intracavity, the heat-transfer thing can be water, oil etc. can realize the material to sunken cavity even heating, also can be the air, so realizes controlling the temperature in the sunken cavity, and then realizes the control to the temperature in the microorganism culture section of thick bamboo.

As a preferable embodiment, a flexible sealing gasket 17 is further arranged on the upper edge of the concave cavity, and the sealing gasket 17 is used for sealing the gap between the outer wall of the culture cylinder and the concave cavity. The provision of the gasket 17 ensures good heat transfer between the cavity and the barrel 1, and also improves the stability of the barrel 1.

In a preferred embodiment, the gasket 17 is a rubber gasket 17.

In a preferred embodiment, the temperature-controlled cabin 15 is provided with a plurality of concave cavities, and each concave cavity is provided with one culture cylinder. Can improve culture efficiency by a wide margin, when cultivateing to the microorganism of the same quantity in addition, can reduce the quantity of microorganism in single barrel 1 by a wide margin, when improving the cultivation quality, still reduced the risk that cross infection appears between the microorganism by a wide margin, also utilize the control to the microorganism sudden change.

In a preferred embodiment, adjacent recessed chambers are spaced apart such that a channel is formed between the culture cartridges for passage of a person after placement of the culture cartridges in the recessed chambers. The staff is convenient to monitor and operate each culture cylinder.

In a preferred embodiment, the microorganism culture system further comprises a box 18, the temperature control cabin 15 and the culture cylinder are arranged in the box 18, and the temperature control cabin 15 is detachably connected with the box 18. Through setting up box 18, improve the wholeness of culture system, when reducing the contamination that the culture section of thick bamboo receives, still convenient transportation.

In a preferred embodiment, the container 18 is a shipping container.

In a preferred embodiment, a gas chamber 19 is further provided in the box 18, the gas chamber 19 is used for storing gas suitable for the microorganism to be cultured, and a gas-filling pipeline adapted to the through hole 10 of the culture cylinder push rod 9 is further provided on the gas chamber 19. The gas chamber 19 has a closed cavity within which new unused gas is stored. The gas filling pipeline is also provided with a gas filling nozzle matched with the pushing through hole 10, and the gas filling nozzle is in separable sealing fit with the push rod 9. After the air in the gas cavity 4 in the culture cylinder is exhausted, the gas cavity 4 is supplemented with new suitable gas suitable for the growth and development of microorganisms through the matching of the gas-adding pipeline and the through hole 10 of the push rod 9.

In a preferred embodiment, the gas chamber 19 is disposed at the top of the tank 18.

In a preferred embodiment, the gas chamber 19 includes a lower plate, an edge of the lower plate is in sliding sealing fit with a side wall of the box 18, a gas storage space of the gas chamber 19 is formed between the lower plate and a top plate of the box 18, the gas supply pipe is disposed on the lower plate and is communicated with the gas storage space of the gas chamber 19, and a driving device for driving the lower plate to move vertically is further disposed in the box 18. In the present application, the driving device is a driving motor, the driving motor drives the lower side plate to move upwards, and the gas in the gas chamber 19 is pressed into the gas cavity 4 of the culture cylinder, so that the suitable gas is supplemented.

In a preferred embodiment, an exhaust gas chamber 20 is further provided in the housing 18, and the exhaust gas chamber 20 communicates with the communicating tube 5 of each of the culture cylinders. In response to certain microbial cultures, suitable gases within the gas chamber 4 may cause serious pollution or be toxic or harmful, and in response to such toxic or harmful gases, in the present embodiment, the canister waste chamber 20 is collected and then treated for decontamination.

Example 4, as shown in figures 1 and 2:

after obtaining the microbial strains, the strains are cultured in the microbial culture system, then the microbial culture system with the cultured microbial strains is transported to a use site, and then the microbes in the culture system are taken out for use on the site.

As a preferred embodiment, after obtaining the microbial species, before culturing the species in the microbial culture system, the microbial culture system is subjected to a gas replacement and culture substrate supplementation time node experiment,

the microorganism culture system gas replacement and culture substrate supplement time node experiment comprises the following steps: setting the temperature control cabin 15 to be suitable for the temperature of growth, development and propagation of microorganisms, placing a culture substrate and strains into a culture substrate cavity 3 of the culture cylinder according to the filling amount of the culture substrate and the placing amount of the microorganism strains in the culture cylinder in the actual transportation process, then filling suitable gas into a gas cavity 4 of the culture cylinder, starting timing, recording a time reading h0, selecting a microorganism mutation rate threshold value and a death rate threshold value according to the requirements of actual microorganism varieties and application fields, and then carrying out the following steps:

and step 3: the amount of the supplementary ingredient d1 is added to the culture substrate chamber 3, and then the gas in the gas chamber 4 is replaced with a new suitable gas,

step 6: the amount of the supplementary ingredient d2 is added to the culture substrate chamber 3, and then the gas in the gas chamber 4 is replaced with a new suitable gas,

when the strain is cultured in a microorganism culture system, according to the time h1-hn, when the time reaches a certain hx, the corresponding component amount dx is supplemented into the culture substrate cavity 3 of the culture cylinder, and meanwhile, the gas in the gas cavity 4 of the culture cylinder is replaced by new suitable gas, wherein x is more than or equal to 1 and less than or equal to n, and x is an integer.

In a preferred embodiment, the transport time is estimated prior to the transport of the microorganism culture system, the appropriate gas volume and the amount of the additional components required prior to transport to the site of use are calculated based on the transport time and the number of culture cylinders in the microorganism culture system, and the appropriate gas volume and the amount of the additional components required are disposed in the housing 18 of the microorganism culture system. In this embodiment, the desired appropriate gas volume and the amount of components of the culture substrate are estimated by estimating the incubation time of the microorganism in the microorganism culture system and then by combining the incubation time with experimental data, which will not be further described in the present application since it is possible for those skilled in the art to derive this without any doubt by routine knowledge.

In a preferred embodiment, the amounts of the medium-supplemented components required for the culture system are grouped according to the actual data h, the amounts of the medium-supplemented components required for the entire microorganism culture system are assigned to the amounts of the medium-supplemented components required for each group at different points in time, and the amounts of the medium-supplemented components are further divided into equal groups according to the number of culture tubes and assigned to the supplemented component groups corresponding to the culture tubes. In practical use, corresponding groups are selected at different time nodes, and then a culture medium adding component of a certain group of components is added into the culture medium cavity 3 of the culture cylinder, so that the accuracy of the adding component is ensured in the aspect of operation process and good quantification.

In a preferred embodiment, the microorganism in the culture cylinder is mixed with the culture substrate and mixed with the product after the culture system is transported to the site of use. The mode makes the culture medium more fully utilized, and particularly can form good microbial environment buffer, and avoid too much adverse effect on the growth and the development and the propagation of the microorganisms due to too large environmental difference.

The above embodiments are only used for illustrating the invention and not for limiting the technical solutions described in the invention, and although the present invention has been described in detail in the present specification with reference to the above embodiments, the present invention is not limited to the above embodiments, and therefore, any modification or equivalent replacement of the present invention is made; all such modifications and variations are intended to be included herein within the scope of this disclosure and the appended claims.

Claims

1. A culture cylinder for microbial cultivation, characterized in that: including the lower extreme seal the open barrel in upper end and with the open tip matched with lid in barrel upper end the barrel inner space divides into culture medium chamber and gas chamber from bottom to top, the culture medium chamber is used for holding the culture medium that suitable microorganism grows, the gas chamber is used for holding the suitable gas that suitable microorganism grows still be provided with gaseous replacement device in the barrel, gaseous replacement device is used for making the gas in the gas chamber is replaced for new suitable gas.

2. The culture cylinder of claim 1, wherein: the gas replacement device comprises a replacement component and a gas discharge component, the gas discharge component is used for communicating the gas cavity with the outer space of the cylinder, and the replacement component is used for pushing gas in the gas cavity to enable the gas in the gas cavity to be discharged out of the cylinder through the gas discharge component.

3. The culture cylinder of claim 2, wherein: the gas discharge assembly comprises a communicating pipe, one end of the communicating pipe is communicated with the gas cavity, and the other end of the communicating pipe penetrates through the barrel body and is communicated with the external space of the barrel body.

4. The culture cylinder of claim 3, wherein: and a stop valve is also arranged on the communicating pipe.

5. The culture cylinder of claim 3, wherein: the communicating pipe is also provided with a one-way conduction device, so that the communicating pipe is in one-way conduction from the inside of the barrel body to the outside of the barrel body.

6. The culture cylinder of claim 5, wherein: the one-way conduction device is arranged at one end, close to the gas cavity, of the communicating pipe.

7. A culture cylinder according to any one of claims 3 to 6, characterized in that: the communicating pipe is vertically arranged along the inner wall of the gas cavity, the lower end of the communicating pipe is in clearance fit with the upper edge of the culture medium cavity, and the upper end of the communicating pipe penetrates through the upper end part of the cylinder body to form an exhaust nozzle.

8. A culture cylinder according to any one of claims 3 to 6, characterized in that: communicating pipe is including wearing out section and vertical section, vertical section is followed the outer wall vertical arrangement of barrel, it is located to wear out section one end in the barrel, with the last edge clearance fit in culture medium chamber, the other end is worn out outside the barrel, with the lower extreme of vertical section links to each other, it is in certainly to wear out the section the barrel is internal to the barrel in the orientation downward sloping outward.

9. A culture system, characterized by: comprising a culture cylinder according to any of claims 1 to 8.

10. A method for culturing microorganisms, comprising: culturing of microorganisms is carried out using a cartridge according to any one of claims 1 to 8.