KR101449144B1 - Rumen Simulation Continuous Culture System - Google Patents

Abstract

The present invention relates to a fermenter comprising an outlet; A lid for opening and closing the fermentation tank; And a discharge pipe including a connection pipe coupled to the discharge port, a downwardly extending lower pipe extending from the connection pipe and a correlation extending upward from the connection pipe; To a ruminal model continuous culture system. According to the present invention, it is possible to provide a rumen model continuous culture system capable of accurately evaluating the rumen digestibility and nutritional value of raw material feed, and also capable of evaluating methane gas generation amount.

Description

{Rumen Simulation Continuous Culture System}

The present invention relates to a ruminal model continuous culture system, and more particularly, to a ruminal model continuous culture system capable of compensating for the disadvantages of the conventional ruminal model continuous culture system and measuring the amount of methane generated in the rumen.

Various methods have been used to evaluate the digestibility and nutritional value of feedstuffs in ruminants. First, the in vivo method is relatively accurate because it directly measures the nutrient digestion of feed in animals, but it takes time and money (Van Straalen and Tamminga, 1990). On the other hand, the in situ method is a relatively simple method in which the nylon bag containing the substrate is cultured in the rumen for a certain period of time and then the amount of the substrate that has escaped from the nylon bag is measured. (Varvikko and Lindberg, 1985). And the in vitro method can overcome limitations of animal biology experiments and is widely used because of its limited time and cost.

Methods for measuring the rate of raw feed degradation in vitro include Tilley and Terry (1963), which measures the amount of gas remaining after a certain period of incubation in ruminal fluid, and the method of measuring and evaluating the amount of gas generated during culture (Menke and Steingass , 1988). However, in order to evaluate in vitro the digestibility and nutritional value of feedstuffs in rumen, a fermentation system should be constructed that meets the actual rumen conditions.

Generally, many batch cultures are performed when culturing in vitro. Although this method can assess the substrate digestibility or the effect of additive under limited conditions, the effect from the in vitro batch culture may not appear in real animals, and the methane production may be different from in vivo. For example, as the proportion of concentrated feed stuff increases in forage to concentrate ratio, methane production decreases in actual rumen but increases in in vitro batch culture. It is necessary to construct an in vitro rumen fermentation system that can realize actual rumen conditions to overcome these problems and to evaluate actual nutrient value.

Unlike in vitro batch culture, the continuous culture method is a culture method in which the substrate or buffer is continuously injected and the culture medium or the digested substrate is also discharged at a constant rate to maintain the incubator environment constant like a rumen.

The Rumen simulation technique (RUSITEC, 1977) proposed by Czerkawski and Breckenridge exists as a conventional continuous culture method.

However, the RUSITEC system has the problem that the fermenter must be opened every time the substrate is replaced through the nylon bag, and the turnover time of the substrate is fixed, so that there is a problem in that it is not possible to predict the parameters that are generated when testing different raw materials .

Later, an improved Rumen Simulation Continuous Culture System (RSCC, 1988) from the RUSITEC system was proposed by Teather and Sauer. (See Fig. 1)

Referring to FIG. 1, a conventional RSCC system includes a T-shaped discharge pipe 20 in a fermentation tank 10 to discharge a culture liquid and feed. At this time, by forming the hole 30 in the portion of the discharge tube 20 which is submerged in the culture liquid, the culture liquid and the feed can be introduced into the discharge tube 20 through the hole 30.

However, in the conventional RSCC system, the dry matter content in the fermentation tank 10 is increased as the experiment period progresses, so that the result is much different from the experimental result of the actual animal, and the discharge tube 20 is frequently clogged to the fermentation tank 10 Had a problem that affected them.

It is an object of the present invention, which has been devised to solve the above-mentioned problems, to provide a rumen model continuous culture system capable of accurately evaluating rumen digestibility and nutritive value of a raw material feed, and also capable of evaluating methane gas generation amount.

According to an aspect of the present invention, there is provided a fermenter comprising: a fermenter having a discharge port; a lid for opening and closing the fermenter; a connection pipe connected to the discharge port; And a discharge pipe including a lower pipe extending downward from the correlation.

Further, the correlation is formed so as to extend in the upper diagonal direction, and the lower tube extends in the lower diagonal direction.

The apparatus further includes a cap inserted into the correlation.

Further, the lid is provided with a stirring rod input port, a feed port, and a buffer input port.

The stirring rod may be inserted into the fermenter through the stirring rod inlet and an agitator for controlling the rotation of the stirring rod.

Further, the stirrer is characterized in that the rotation speed of the stirring rod is controlled to 10 to 20 rpm.

The apparatus further includes a feed inlet pipe inserted into the fermentation tank through the feed inlet and a feed inlet rod inserted into the feed inlet pipe.

The apparatus also includes a peristaltic pump for supplying a buffer container for receiving the buffer and a buffer accommodated in the buffer container to the fermenter through the buffer inlet.

The peristaltic pump is characterized in that the buffer is supplied in an amount of 1000 to 1200 mL / day.

The apparatus also includes a Tedlar bag filled with gas and connected to the buffer container.

Further, the gas filled in the Tedlar bag is carbon dioxide.

Further, the lid is further provided with an electrode input port.

The apparatus further includes an electrode that is injected into the fermentation tank through the electrode inlet, and a pH meter that measures pH in the fermentation tank through the electrode.

Further, the lid is further provided with a gas outlet.

The apparatus may further include a gas collecting pipe connected to the gas discharging port and a gas collecting bag collecting gas discharged through the gas collecting pipe.

The apparatus further includes a methane gas meter for measuring the amount of methane gas captured in the gas collection bag.

The apparatus further includes a circulation water tank for surrounding the fermentation tank, and a hot water circulator for keeping the temperature of the circulation water tank constant.

The hot water circulator is characterized in that the temperature of the circulating water tank is maintained at 37 to 40 캜.

As described above, according to the present invention, it is possible to provide a continuous rumen model continuous culture system capable of easily measuring the fermentation gas and the like without an experimental animal in a laboratory.

Further, according to the present invention, it is possible to provide a rumen model continuous culture system having a discharge tube without clogging.

1 is a diagram illustrating a conventional RSCC system.
2 is a view showing a fermenter and a discharge pipe according to an embodiment of the present invention.
3 is a view illustrating a lid according to an embodiment of the present invention.
4 is a view showing a stirring rod and a stirrer according to an embodiment of the present invention.
5 is a view showing a feed inlet pipe and a feed rod according to an embodiment of the present invention.
FIG. 6 is a view showing the state where the feed inlet pipe and the feed input rod shown in FIG. 5 are engaged.
7 is a view showing a buffer container and a peristaltic pump according to an embodiment of the present invention.
8 is a view showing an electrode and a pH meter according to an embodiment of the present invention.
9 is a view showing a gas collecting tube and a gas collecting bag according to an embodiment of the present invention.
10 is a view showing a circulating water tank and a hot water circulator according to an embodiment of the present invention.

The details of other embodiments are included in the detailed description and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims. Like reference numerals refer to like elements throughout the specification.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a ruminal model continuous culture system according to an embodiment of the present invention will be described with reference to embodiments of the present invention and drawings for explaining the same.

2 is a view showing a fermenter and a discharge pipe according to an embodiment of the present invention.

Referring to FIG. 2, the rumen model continuous culture system according to an embodiment of the present invention may include a fermenter 100 and a discharge tube 200.

The fermenter (100) has a predetermined receiving space (130) for receiving a culture liquid and a substrate therein.

In addition, the fermentation tank 100 may include a discharge port 110 so that the culture liquid or the like can be discharged to the outside during the culturing.

At this time, the discharge port 110 may be formed on the outer peripheral surface of the fermentation tank 100 and may be combined with the discharge pipe 200.

The fermentation tank 100 is preferably formed of a glass material, and may be formed of, for example, pyrex glass.

In addition, the capacity of the fermentation tank 100 can be variously changed. In a subsequent experiment, a 2 L fermenter (100) was used, and the culture liquid contained in the fermenter (100) was set to 1 L.

The temperature of the fermentation tank 100 may be kept constant by a water jacket method.

The discharge tube 200 is coupled with the discharge port 110 to discharge the culture liquid and the like contained in the fermentation tank 100 to the outside.

The discharge pipe 200 includes a connection pipe 210 coupled to the discharge port 110, a correlation 220 extending upward from the connection pipe 210 and a lower pipe 230 extending downward from the correlation pipe 220 Lt; / RTI >

The connection pipe 210 is connected to the discharge port 110 of the discharge pipe 200 and may be coupled to the discharge port 110 through a separate connection member 150.

The correlation 220 may extend upward from the coupling pipe 210. Specifically, the correlation 220 is preferably extended toward the upper diagonal direction as shown in FIG. 2 in order to minimize clogging of the discharge tube 200.

At this time, the angle [theta] of the correlation 220 may be variously set, but it is preferably set to about 30 to 60 degrees with respect to the ground. Then, in the experiment, the angle? Of the correlation 220 was set to 45 degrees.

The lower tube 230 may extend from the correlation 220 toward the lower side. Specifically, the lower pipe 230 may be extended toward the lower diagonal direction as shown in FIG.

Therefore, a culture solution or the like in the fermentation tank 100 can be discharged to the outside through the discharge port 110, the connection pipe 210, and the lower pipe 230.

Therefore, by using the above-described 'V' shaped discharge tube 200 without using the conventional 'T' shaped discharge tube 20, clogging of the discharge tube 200 can be minimized during the discharge of the culture liquid .

At this time, a cap 250 to be inserted into the correlation 220 may be additionally provided. The stopper 250 may also be movable within the correlation 220.

When the stopper 250 is inserted deeply into the connection pipe 210 beyond the lower pipe 230, the stopper 250 closes the lower pipe 230, so that the discharge of the culture liquid or the like can be blocked. In addition, when the discharge tube 200 is clogged, clogging may be eliminated through the clog 250.

3 is a view illustrating a lid according to an embodiment of the present invention.

Referring to FIG. 3, the lid 300 according to an embodiment of the present invention may include a stir rod inlet 310, a feed inlet 320, and a buffer inlet 330.

The lid 300 serves to open and close the fermentation tank 100 and can be coupled to the upper side of the fermentation tank 100.

In addition, the lid 300 may further include an electrode inlet port 340 and / or a gas outlet port 350.

The stirring rod input port 310 is preferably formed at the center of the lid 300 and other feed inlet port 320 and buffer inlet port 330 may be formed around the stirring rod inlet port 310.

The lid 300 is preferably formed of the same material as the fermentation tank 100.

4 is a view showing a stirring rod and a stirrer according to an embodiment of the present invention. 4, the stirring rod 410 and the stirrer 420 are mainly shown for convenience of explanation.

Referring to FIG. 4, the rumen model continuous culture system according to an embodiment of the present invention may further include a stirring rod 410 and a stirrer 420.

The stir rod 410 may be inserted into the fermenter 100 through the stirring rod inlet 310 formed in the lid 300. At this time, the stirring rod 410 can be controlled by the stirrer 420.

The stirring rod 410 may be formed of various materials, for example, stainless steel.

A bar-shaped paddle 411 may be formed at a lower end of the stirring rod 410 to improve the agitating force, and a spiral plate 412 may be provided at a predetermined distance from the upper side of the paddle 411 .

The paddle 411 existing in the stirring rod 410 may be positioned to be spaced apart from the bottom of the fermenter 100.

In a later experiment, the paddle 411 was positioned at a distance of about 5 mm from the bottom of the fermentation tank 100, and the spiral plate 412 was positioned at a height similar to the discharge port 110.

The stirrer 420 is connected to the stirring rod 410 to control the rotation of the stirring rod 410, the rotation speed, and the like.

At this time, the stirrer 420 can control the rotation speed of the stirring rod 410 to 10 to 20 rpm when culturing.

FIG. 5 is a view showing a feed inlet tube and a feed rod according to an embodiment of the present invention, and FIG. 6 is a view showing a state of engagement between the feed rod and the feed rod shown in FIG. 6, the feed inlet pipe 510 and the feed inlet rod 520 are mainly shown for convenience of explanation.

Referring to FIG. 5, the rumen model continuous culture system according to an embodiment of the present invention may further include a feed inlet pipe 510 and a feed inlet rod 520.

The feed inlet pipe 510 is vertically elongated and can be inserted into the fermenter 100 through a feed inlet 320 formed in the lid 300.

At this time, it is preferable that the feed inlet pipe 510 is positioned so that its lower end portion is submerged in gastric juice.

Also, care should be taken that the feed inlet pipe 510 does not contact the stirring rod 410.

The feed input rod 520 may be inserted into the feed inlet pipe 510 to close the feed inlet pipe 510. In addition, the feed input pipe 510 may be used to push the fed feed.

It is preferable to seal the connection portion between the feed inlet pipe 510 and the feed inlet port 320 using Teflon tape or the like in order to block the air inside the fermentation tank 100 during the culturing. It is also desirable to seal the connecting portion between the feed inlet pipe 510 and the feed inlet rod 520.

Thus, the substrate for the experiment can be fed through the feed inlet tube 510. In later experiments, the substrate feed was set at 10 g / day (day) and the turn rate of the substrate was set at 0.017 h ^-1 .

7 is a view showing a buffer container and a peristaltic pump according to an embodiment of the present invention. In particular, in FIG. 7, the buffer container 620 and the peristaltic pump 630 are mainly shown for convenience of explanation.

Referring to FIG. 7, the ruminal model continuous culture system according to the embodiment of the present invention may further include a buffer container 620 and a peristaltic pump 630.

The buffer container 620 is a container for receiving the buffer 610 therein, for example, a triangular flask or the like can be used.

The peristaltic pump 630 serves to supply the buffer 610 accommodated in the buffer container 620 into the fermenter 100 through the buffer inlet 330.

The peristaltic pump 630 may include a first tube 661 connected to the buffer vessel 620 and a second tube 662 connected to the buffer inlet 330. [

The buffer 610 in the buffer vessel 620 may be introduced into the peristaltic pump 630 through the first tube 661 and the peristaltic pump 630 may be supplied with the buffer 610 corresponding to the pre- 2 tube 662 and the buffer inlet 330 to the fermenter 100.

The peristaltic pump 630 preferably controls the buffer supply amount in an amount of 1000 to 1200 mL / day (day).

At this time, it is preferable to connect a tedlar bag 650 filled with the gas to the buffer container 620 so that the internal pressure of the buffer container 620 containing the buffer 610 can be maintained .

At this time, the Teddler bag 650 may be connected to the buffer container 620 through a separate tube 663.

Carbon dioxide (CO ₂ ) or the like may be used as the gas to be filled in the Tedlar bag 650.

8 is a view showing an electrode and a pH meter according to an embodiment of the present invention. In particular, the electrode 710 and the pH meter 720 are mainly shown in FIG. 8 for convenience of explanation.

8, the ruminal model continuous culture system according to the embodiment of the present invention may further include an electrode 710 and a pH meter 720 for measuring the hydrogen ion index in the fermentation tank 100.

The electrode 710 may be inserted into the fermenter 100 through the electrode inlet 340 formed in the lid 300.

At this time, it is preferable that the electrode 710 has a sufficient length to be submerged in the culture liquid of the fermentation tank 100. Care should be taken that the electrode 710 does not touch the stirring rod 410 located in the fermenter 100.

The pH meter 720 may be connected to the electrode 710 to measure the pH of the fermentation tank 100 through the electrode 710.

The pH meter 720 can measure the temperature as well as the pH value in the fermentation tank 100 via the electrode 710.

9 is a view showing a gas collecting tube and a gas collecting bag according to an embodiment of the present invention. 9, the gas collecting pipe 810, the gas collecting bag 820 and the methane gas analyzer 830 are mainly shown for convenience of explanation.

Referring to FIG. 9, the rumen model continuous culture system according to the embodiment of the present invention may further include a gas collection pipe 810 and a gas collection bag 820.

The gas collecting pipe 810 can connect between the gas discharging port 350 formed in the lid 300 and the gas collecting bag 820.

Accordingly, the gas generated in the fermentation tank 100 can be collected in the gas collection bag 820 through the gas collection pipe 810.

At this time, as the gas collection bag 820, a tedlar bag can be used.

Also, a methane gas measuring instrument 830 for measuring the methane gas amount in the gas collected in the gas collecting bag 820 may be provided.

Therefore, the ruminal model continuous culture system according to the embodiment of the present invention can measure the methane gas generated in the rumen.

10 is a view showing a circulating water tank and a hot water circulator according to an embodiment of the present invention. In particular, in FIG. 10, circulation water tank 910 and hot water circulator 920 are mainly shown for convenience of explanation.

As shown in FIG. 10, the fermenter 100 may be located in the circulation water tank 910.

Therefore, the temperature of the fermentation tank 100 can be kept constant by the circulation water tank 910 surrounding the fermentation tank 100.

The hot water circulator 920 circulates hot water through the first connection port 931 and the second connection port 932 formed in the circulation water tank 910. To this end, each of the connectors 931 and 932 and the hot water circulator 920 may be connected to the tube 940.

At this time, it is preferable that the temperature of the circulating water tank 910 is maintained at 37 to 40 ° C.

Hereinafter, an experimental method and an experimental result using the rumen model continuous culture system according to an embodiment of the present invention will be described.

First, a method of manufacturing a buffer to be used in an experiment will be described. (1L standard)

Add 250 mL of distilled water to a 2 L Erlenmeyer flask. Add 0.12 mL of micromineral solution and stir. Add 250 mL of distilled water to the Erlenmeyer flask and stir for 20 minutes.

Add 250 mL of in vitro buffer solution and stir for 10 minutes. Add 250 mL of a macromineral solution and stir for 15 minutes.

1.25 mL of resazurin solution is added and stirred. The inlet of the Erlenmeyer flask is covered with aluminum foil and heated by bubbling the carbon dioxide (CO 2) gas from the anaerobic digester. When the solution starts to boil, heat it for about 10 minutes and stop. Add 50 mL of the reduction solution to the cooled solution. When the color of the solution changes to colorless, the buffer preparation is completed.

Hereinafter, an experimental method will be described.

The culture medium is obtained by the following procedure. First, the collected gastric juice is filtered with 8 layers of gauze and then filtered with glass wool. The stomach fluid is kept anaerobic by bubbling with CO2 gas. Pre-prepared buffer is mixed with gastric juice (250 mL of gastric juice, 600 mL of buffer) while maintaining the carbon dioxide gas.

The fermenter 100 is charged with 10 g of a substrate. Further, the buffer 610 is put into the buffer vessel 620 while maintaining the anaerobic state. The Teddler bag 650 filled with the carbon dioxide gas is connected to the buffer vessel 620 and the tube connected to the buffer vessel 620 is connected to the buffer inlet 330 of the lid 300.

The gas collecting bag 820 is connected to the gas outlet 350 of the lid 300 using the gas collecting pipe 810. As the gas collecting tube 810, a gas tube may be used.

The electrode 710 is inserted through the electrode input port 340 of the lid 300 so as to sufficiently contact the culture liquid. At this time, care is taken that the electrode 710 does not contact the stirring rod 410.

The feed inlet pipe 510 is inserted through the feed inlet 320 of the lid 300 so as to be sufficiently immersed in the culture liquid.

The lid 300 is coupled with the fermentation tank 100, and the anaerobic state is maintained by using a vacuum grease and a Teflon tape.

Carbon dioxide gas is introduced into the fermentation tank 100 through the gas outlet 350 of the lid 300 for about 10 minutes. (The inside of the fermenter 100 is changed to the anaerobic state, and the carbon dioxide gas is continuously flowed until the start of cultivation)

Seal all connection points with vacuum grease and Teflon tape.

The peristaltic pump 630 is operated to input the buffer 610 into the fermenter 100.

The culture medium in the anaerobic state is introduced into the fermentation tank 100 through the feed inlet 320. (The culture solution is poured until the culture liquid overflows through the discharge port 110)

After that, stop the injection of carbon dioxide gas and block all the inlets. Further, the circulation water tank 910 and the fermentation tank 100 are connected and the temperature is set (39 DEG C). The rpm of the stirring rod 410 is set, and the culture is started.

The experimental results are summarized as follows.

(A) pH: The pH of fermentation broth was constantly maintained at 6.8 ~ 7.1 during the whole experimental period and decreased after feeding.

(B) Temperature: The temperature of the fermenter was kept constant at 39 ~ 40 ℃ during the whole experimental period, and it tended to increase after feeding.

(C) Dry matter content of the culture and overflow: During the test run of the rumen model continuous culture system, the content (% AF) of the discharged samples collected through the fermenter and the discharge pipe (200) The dry matter content of the fermenter was 1.77 (± 0.437)% and the dry matter content of the discharged sample was 1.77 (± 0.423)%. It can be seen that the mixing of the culture medium is performed well by relatively stirring (10 to 20 rpm), and the ejection speed of the solid phase and the liquid phase is maintained to be similar. The dry matter content of the discharged samples tended to increase after feeding, while the dry matter content of the fermenter was not influenced by feed input. In addition, the dry matter content of the fermenter tended to be maintained at a constant level as culture continued. The decrease in the dry matter content of the culture medium is believed to be due to the fast keeping of the buffering rate during the measured period of the dry matter content.

(D) Gas production rate: The amount of gas production per hour was 29.07 (± 25.52) mL on average during the whole test operation period, showing a relatively large variation. No effect on feeding was found.

(E) Methane emission: The ratio of methane in the collected gas over a total of four times during the test period of the rumen continuous culture system was measured and it was found that it occupied an average of 7.08 (± 0.796)% of the generated gases. No particular trends were observed with feed input.

(F) Results of fermentation

Ⓛ Fermentation characteristics: The pH can be continuously measured and recorded to confirm the change of pH. The pH tends to increase rapidly after the substrate is supplied. The apparent digestibility and true digestibility of the substrate were 42.8% and 61.6%, respectively, and the average amount of total volatile fatty acids (VFA) was 51.4 mmol / d. The concentration of volatile fatty acids was diluted in the fermentation system and was about 1/3 of that in the rumen. In the composition of volatile fatty acids, the proportion of butyrate increased during the adaptation period, but the composition of volatile fatty acids after the adaptation period did not differ from that of the actual ruminant. Cellulase activity was similar to that of ruminant juice, but xylanase activity was higher in the culture system than in ruminant.

② Microbial community: Protozoa decreased more than ruminant, but Holotrich decreased in protozoa. Total rRNA was also reduced compared to ruminal juice, but there was no difference in the proportion of bacteria or eukaryotes (protozoa and fungi) in rRNA, and the ratio of archaea was two times higher than that of ruminant juice. The rRNA concentration of cellulolytic microorganisms (CMO) was lower than the ruminal fluid, but the ratio of CMO to total rRNA was not different. The proportion of Fibrobacter was maintained in the fibrinolytic microorganism. The proportion of Chytridiomycetes and R. albus was increased more than that of ruminants and the ratio of R. flavefaciens was decreased.

To summarize, the ruminal model continuous culture system used in this experiment has the following differences from the existing system.

First, a gas outlet 350 is installed in the lid 300 to collect fermentation gas through fermentation of the system, and in particular, methane gas generated in the rumen can be measured.

In addition, since a discharge vessel 200 and a cap 250 are installed at the discharge port 110 of the fermentation tank 100, it is possible to prevent clogging at the discharge port compared with the conventional system by periodic rotation, . In addition, the use of a computer program enables precise control of the experimental environment.

It will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. The scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning and scope of the claims and the equivalents thereof are included in the scope of the present invention Should be interpreted.

100: fermenter 110: outlet
200: discharge pipe 210: connection pipe
220: correlation 230:
250: cap 300: cap
310: Stirring rod inlet port 320: Feed inlet port
330: buffer input port 340: electrode input port
350: gas outlet 410: stir bar
420: Stirrer 510: feed feed pipe
520 Feed feed rod 610 Buffer
620: buffer vessel 630: peristaltic pump
650: First Teddler bag 710: Electrode
720: pH meter 810: Gas collecting tube
820: Gas collection bag 830: Methane gas measuring instrument
910: Circulating water tank 920: Hot water circulator

Claims (18)

A fermentation tank having an upper opening and an outlet at an outer periphery;
A lid coupled to the upper side of the fermentation tank; And
A discharge pipe including a connection pipe connected to the discharge port, a correlation pipe extending upward from the connection pipe, and a lower pipe extending downward from the correlation pipe; Lt; RTI ID = 0.0 > ruminal < / RTI > continuous culture system. The method according to claim 1,
The correlation is formed in the upper diagonal direction,
Wherein the lower tube extends in a lower diagonal direction. The method according to claim 1,
A plug inserted into the correlation; A ruminal model continuous culture system. The lid according to claim 1,
A stirring rod inlet, a feed inlet, and a buffer inlet. 5. The method of claim 4,
A stirring rod inserted into the fermenter through the stirring rod inlet; And
An agitator for controlling the rotation of the stirring rod; A ruminal model continuous culture system. 6. The agitator according to claim 5,
Wherein the stirring speed of the stirring rod is controlled at 10 to 20 rpm. 5. The method of claim 4,
A feed inlet pipe inserted into the fermenter through the feed inlet; And
A feed inlet rod inserted into the feed inlet pipe; A ruminal model continuous culture system. 5. The method of claim 4,
A buffer container for receiving a buffer; And
A peristaltic pump for feeding the buffer contained in the buffer vessel into the fermenter through the buffer inlet; A ruminal model continuous culture system. 9. The peristaltic pump according to claim 8,
Wherein the buffer is supplied in an amount of 1000 to 1200 mL / day. 9. The method of claim 8,
A Tedlar bag filled with a gas and connected to the buffer container; A ruminal model continuous culture system. 11. The method of claim 10, wherein the gas filled in the Tedlar bag
Wherein the ruminal model continuous culture system is carbon dioxide. 5. The lid according to claim 4,
And a further electrode inlet is formed in the rumen model continuous culture system. 13. The method of claim 12,
An electrode charged into the fermenter through the electrode inlet; And
A pH meter for measuring pH in the fermentation tank through the electrode; A ruminal model continuous culture system. 5. The lid according to claim 4,
Wherein the gas outlet is further formed. 15. The method of claim 14,
A gas collection pipe connected to the gas discharge port; And
A gas collecting bag collecting the gas discharged through the gas collecting pipe; A ruminal model continuous culture system. 16. The method of claim 15,
A methane gas measuring device for measuring an amount of methane gas captured in the gas collecting bag; A ruminal model continuous culture system. The method according to claim 1,
A circulation water tank for surrounding the fermentation tank; And
A hot water circulator for keeping the temperature of the circulation water tank constant; A ruminal model continuous culture system. 18. The hot water circulating system according to claim 17,
Wherein the temperature of the circulation water bath is maintained at 37 to 40 캜.

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