KR101808957B1 - Feed additive for reducing methane emission from ruminants comprising sinigrin - Google Patents

Abstract

The present invention relates to a feed additive composition for reducing methane production, and more particularly, to a feed additive composition for reducing methane production of a ruminant including Sinigan and a method for inhibiting methane production of a ruminant using the feed additive composition.
According to the present invention, when the shinigin is treated at a specific concentration, only the methane production can be significantly reduced without affecting the pH, the digestibility of the digestion, and the like, It is expected that it can be usefully used as a feed additive or a feed composition for reducing the amount of the feed.
In addition, according to the present invention, it is possible to reduce the energy loss to the ruminant methane by effectively reducing the methane-reducing effect while minimizing the physiological side effects of the ruminant due to the addition of a large amount of glucaninolate, Increased energy can also be helpful for productivity.

Description

[0001] The present invention relates to a feed additive for reducing methane production from ruminants comprising sinryrin,

The present invention relates to a feed additive composition for reducing methane production, and more particularly, to a feed additive composition for reducing methane production of a ruminant including Sinigan and a method for inhibiting methane production of a ruminant using the feed additive composition.

Global warming is accelerating due to the increase in global warming gas, resulting in extreme weather, crop production and food shortages. Global warming is a phenomenon in which the average temperature on the surface of the earth rises. It is known that many greenhouse gases enter the atmosphere and are absorbed or re-emitted by infrared radiation. The IPCC (Intergovernmental Panel on Climate Change) CO _2), methane (CH _4), nitrous oxide (N ₂ O), hydrogen fluorocarbons (HFCs), perfluorocarbons (PFCs) and sulfur hexafluoride (SF ₆₎ was selected as the six greenhouse gases. Due to the increase in greenhouse gas concentration due to overuse of fossil fuels after industrialization, the temperature of the earth is continuously rising.

Dual methane (CH ₄₎ gas is known to be due to the influence on the global warming only less amount of carbon dioxide that is produced as compared to 21 times higher, cause the carbon dioxide and then with a significant impact on global warming. Such methane gas is produced by fermentation of microorganisms such as ponds, wetlands, rice fields, fermentation of intestines of ruminant livestock and decomposition of manure, or by natural and industrial activities such as coal, oil, natural gas, and exhaust gas. In particular, methane is generated during the normal digestion process (intestinal fermentation) on cattle, water buffalo, sheep, goats, and camels. The methane gas emitted by the livestock sector now accounts for 13 to 19% Respectively.

Therefore, there is a desperate need for research to improve the productivity of animals and to preserve the global environment by minimizing methane production in rumen.

For this purpose, a method using chemicals such as halogen compounds, ionophore antibiotics, 2-bromoethanesulphonate, and lumazine has been mainly used. However, the methods proposed so far have not only a small effect of reducing the methane generation, but also have a problem in that the methane inhibition effect varies depending on the composition of the feed. In addition, there has been a problem that the ruminal microorganism adapts to the substance or degrades the substance when it is treated for a long time, and the effect is not continuous.

Therefore, it is urgent to develop effective substances for increasing the productivity of ruminants and reducing methane production in ruminants.

DISCLOSURE Technical Problem The present invention has been made in order to solve the above-mentioned problems in the prior art. The present inventors have made efforts to increase the productivity of ruminants and to reduce the generation of methane in ruminants, sinigrin) has an excellent effect on the reduction of methane production in rumen without affecting pH and dry digestibility of rumen. Based on this finding, the present invention has been completed.

Accordingly, an object of the present invention is to provide a feed additive composition for reducing methane production of ruminants, which comprises sinigrin as an active ingredient.

Another object of the present invention is to provide a feed composition for reducing methane production of a ruminant compounded with the above additive composition.

Another object of the present invention is to provide a method for inhibiting the methanogenesis of ruminants comprising feeding sinigrin to an animal.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention provides a feed additive composition for reducing methane production of ruminants, which comprises sinigrin as an active ingredient.

The present invention provides a feed composition for reducing methane production of a ruminant compounded with the feed additive composition.

In one embodiment of the present invention, the syngene can be formulated at a concentration of 10 to 130 탆 ol.

The present invention provides a method for inhibiting methane production in a ruminant, comprising feeding sinigrin to the animal.

In one embodiment of the present invention, the syngene can be fed at a concentration of 10 to 130 μmol.

According to the present invention, when the shinigin is treated at a specific concentration, only the methane production can be significantly reduced without affecting the pH, the digestibility of the digestion, and the like, It is expected that it can be usefully used as a feed additive or a feed composition for reducing the amount of the feed.

In addition, according to the present invention, treatment with a specific concentration of glucurinolate, cininogue, is effective in reducing methane while minimizing the occurrence of physiological side effects of ruminants due to the addition of a large amount. Therefore, reducing the energy lost to methane by ruminants has the added benefit of being more productive because of the increased energy available.

FIG. 1 shows the results of confirming the change of pH with incubation time after treatment of cinchinel with concentration (20, 40, 60, 80 μmol).
FIG. 2 shows the results of confirming total gas production change according to the incubation time after treatment with cinnamin (20, 40, 60, 80 μmol).
FIG. 3 shows the results of confirming the change in methane gas production with time after culturing Sinnigreen at various concentrations (20, 40, 60, 80 μmol).
FIG. 4 shows the result of confirming the change of microbial protein synthesis according to the incubation time after treating the cinchinis with concentration (20, 40, 60, 80 μmol).
FIG. 5 shows the results of confirming the change of ammonia nitrogen (NH ₃ -N) concentration according to the incubation time after treatment with cinnamin (20, 40, 60, 80 μmol)
FIG. 6 shows the results of confirming the changes in the digestibility of dry matter according to the incubation time after treatment with cinnamin (20, 40, 60, 80 μmol).
FIG. 7 shows the results of confirming the changes in the amount of volatile fatty acid (VFA) produced during the incubation time after treatment with cinnamin (20, 40, 60, 80 μmol).
FIG. 8 shows the results of confirming the change in the content of cinnamin according to the time of culture using HPLC.
FIG. 9 shows the result of the change in the methane gas production amount after adding the cinnamin to the basic feed in the animal model at the concentration (40, 60, 120 μmol).

Unlike conventional studies in which sinigrin causes adverse effects on the breeding physiology and productivity of ruminants, the present inventors have found that when sinigrin is treated at a certain concentration, it affects ruminal pH and dry digestibility And that the amount of methane produced in the rumen is significantly reduced.

Hereinafter, the present invention will be described in detail.

The present invention provides a feed additive composition for reducing methane production of ruminants, which comprises sinigrin as an active ingredient.

The term "ruminant" in the present invention refers to any mammal of the mammal of Amuminous Ruminantia, which has a characteristic of chewing out the swallowed food once in its digestive form and is divided into three or four chambers called rumen Means an animal. Examples of such ruminants include, but are not limited to, deer, nutrition, buffalo, cattle, sheep, camels, goats, giraffes, and the like.

A sinigrin containing the active ingredient in the composition of the present invention may have the structure of formula (1). Further, it is apparent to those skilled in the art that cinchinens can be obtained from plant extracts and chemically synthesized, but also those chemically synthesized exhibit the same effects as those extracted.

[Chemical Formula 1]

According to one embodiment of the present invention, the total gas production amount and the methane production amount were examined while the cinchinis were treated at a specific concentration through the in vitro rumen fermentation test. As a result, compared to the control without the cinchinen, It was confirmed that the total gas production amount and the methane production amount were significantly decreased in one test section (see Examples 3 and 4).

According to another embodiment of the present invention, it was confirmed that no significant difference was observed between the control and all the test groups as a result of checking the pH change, the ammonia nitrogen, the dry digestibility, and the like while treating the Sinignan with a specific concentration Examples 2, 6 and 7, etc.).

In addition, according to another embodiment of the present invention, it was confirmed that the amount of methane production was significantly reduced in all of the test plants treated with Sinignan in vivo (see Example 10).

Therefore, it has been confirmed not only in vitro but also in vivo that methane production can be significantly reduced without affecting the pH or the digestibility of the digested sample, Once identified, certain concentrations of syngene can be added to the feed composition using the feed as a feed additive to reduce the methane production of the ruminant.

In another aspect of the present invention, there is provided a feed composition for reducing methane production of a ruminant compounded with the feed additive composition, wherein the syngene is preferably formulated at a concentration of 10 to 130 μmol, Preferably, it can be formulated at a concentration of 20 to 80 μmol.

As used herein, the term "feed" means any natural or artificial diet, single meal, or the like component for feeding, ingesting, digesting or suitable for the animal.

The feed composition according to the present invention may include components commonly added to diets such as carbohydrates, proteins, fats, minerals, vitamins, minerals, and water in addition to cinchignan. Any of those commonly used in the art can be used.

In addition, the feed composition according to the present invention can be produced in the form of a conventional feed composition such as powder and pellets, but it is not limited thereto and can be produced in various types of feed known in the art.

In another aspect of the present invention, the present invention provides a method for inhibiting methane production in a ruminant comprising feeding sinigrin to an animal.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

[Example]

Example  1. Experimental Preparation and Experimental Methods

1-1. Prepare microbial inoculum in rumen

In vitro ruminal fermentation test for rumen microbial inoculum preparation was carried out by using eight layers of cheese cloth collected from the rumen in the milk of cannulae. The separated gastric juice was kept at 39 ℃ using a thermos bottle and immediately transported to the laboratory. Then, a CO ₂ gas was continuously injected using a 5L bioreactor equipped with a rubber stopper to maintain the anaerobic condition of the 39 ° C artificial saliva and ruminant The gastric juice was mixed at a ratio of 1: 1. They were subcultured using hay (rice straw, 20g / L) and concentrated feed (20g / L) as a substrate and incubated for 24 hours at 39 ° C in an incubator. The ruminal fluid was centrifuged (2500 rpm × 15 min) and the supernatant was taken and used as the inoculum for microbial inoculation in the rumen.

1-2. In vitro  Rumen fermentation

In vitro culture conditions for microbial fermentation characteristics in rumen were examined by using a serum bottle (200 mL), a control group without shinigreen treatment and a test group treated with shinigreen (20, 40, 60, 80 μmol) (Inoculum) (5 mL) prepared in Example 1-1 was inoculated with a mixture of CO ₂ gas (100 mL) and an experimental saline The rubber stopper and the aluminum lid were sealed with a dispenser as a fixing device. All incubation conditions were maintained while maintaining the anaerobic condition. After the inoculation, the culture was carried out at 39 ° C in an incubator.

Example  2. Shinny Green  Determination of pH change by addition

2, 4, 6, 8, 10, and 10 days after inoculation with the microbial inoculum in the control and test groups according to Example 1-2, using a pH meter (Orion 3 star, Thermo scientific, USA) 12 < / RTI > and 24 hours).

As a result, as shown in Fig. 1, it was confirmed that the pH decreased as the cultivation proceeded, both in the control without the syngene and in the test with the shinigreen. When examined within the range suitable for normal rumen microbial fermentation, no negative effects were observed.

From the above results, it was found that the addition of cinchinel to the rumen juice did not significantly affect the pH in the rumen.

Example  3. Shinny Green  By addition Total gas  Identification of changes in total gas production

2, 4, 6, and 8 using the gas production material (Model Ab-5210, AITEC Inc., Canada) while inoculating and inoculating the microorganism inoculum into the control and test sites according to Example 1-2. , 10, 12 and 24 hours).

As a result, as shown in FIG. 2, it was confirmed that as the incubation time was increased as compared with the control, the total gas production amount of the test strip added with Sinignan decreased. Especially, total gas production was significantly lower in all test groups than in the control group after 6 hours of culture (p <0.05). In addition, the control group showed the highest total production of 146.13 mL (p <0.05) at the 24th day of culture compared to the control group, while the lowest value of 99.80 mL of total gas production (P < 0.05).

Example  4. Shinny Green  Change of methane production by addition

2, 4, 6, 8, 10, 12, and 24 hours) using a syringe connected to a 3-way stopper while inoculating and incubating the microbial inoculum in the control and test sites according to Example 1-2. And the amount of methane produced was measured by gas chromatography (GC).

As a result, as shown in FIG. 3, it was confirmed that the amount of methane produced in the test strip added with cinigne compared to the control was significantly decreased over the entire culture time (p <0.05). In particular, when 80 μmol of shinigreen was treated, it was confirmed that methane production was reduced by 90% or more when cultured for 24 hours compared with the control.

Example  5. Shinny Green  Identification of microbial protein synthesis by addition

2, 4, 6, 8, 10, 12, and 24 hours) while inoculating the microorganism inoculum into the control and test sites according to Example 1-2 and culturing the cells. The cells were harvested by centrifugation (Brushless DC centrifugation (2,500 rpm x 15 min) using a centrifuge (VS-6000CF, Vision Scientific Co., Ltd., Korea). After centrifugation, the supernatant was taken and microbial protein synthesis (MPS) changes were measured according to the method of Lowry et al. (1951).

As a result, as shown in Fig. 4, it was confirmed that the MPS tended to decrease as the incubation time increased in the test group supplemented with Sinignan. (P <0.05), while the control was slightly lower (p <0.05) in the control group treated with 60 umol of shinigin (p <0.05) And there was no significant difference between the two groups.

Example  6. Shinny Green  The addition of ammonia nitrogen (NH ₃ -N) concentration change confirmation

2, 4, 6, 8, 10, 12, and 24 hours) while inoculating the microorganism inoculum into the control and test sites according to Example 1-2 and culturing the cells. The cells were harvested by centrifugation (Brushless DC centrifugation (2,500 rpm x 15 min) using a centrifuge (VS-6000CF, Vision Scientific Co., Ltd., Korea). After centrifugation, the supernatant was measured and the ammonia nitrogen (NH ₃ -N) concentration was measured using a spectrophotometer (Spectroic PC system 4D-5210, Thermo scientific, USA) according to the method of Chaney and MArbach (1962).

As a result, as shown in FIG. 5, it was confirmed that the ammonia nitrogen (NH ₃ -N) concentration tended to decrease as the culture progressed in the control and all the test sites. In particular, the ammonia nitrogen (NH ₃ -N) concentration in the control and all test groups decreased rapidly during 4 ~ 6 hours of incubation, but no significant difference was observed between the control and test groups.

From the above results, it was found that the addition of cinigne to ruminant juice had no significant effect on ammonia nitrogen concentration in the rumen.

Example  7. Shinny Green  Identification of change in digestibility of dry matter by addition

(0, 2, 4, 6, 8, 10, 12, and 24 hours) by using the method of Van Soest et al. (1967) while inoculating and culturing the microbial inoculum into the control and test sites according to Example 1-2 ) Were used to measure the digestibility of the building. On the other hand, the dry matter content of the test feed was dried for 72 hours in a dry oven at 60 ° C just before analysis.

As a result, as shown in FIG. 6, the digestibility of the dry matter increased with the progress of culture in the control and all the test sites, and it was confirmed that there was no significant difference between the control and the test sites.

From the above results, it can be seen that the addition of cinigne to the ruminal juice does not significantly affect the digestibility of the rumen within the rumen.

Example  8. Shinny Green  Addition of volatile fatty acids VFA ) Confirmation of the amount of change

2, 4, 6, 8, 10, 12, and 24 (%) by using gas chromatography (GC) while inoculating and inoculating the microorganism inoculum into the control and test sites according to Example 1-2. Volatile fatty acids (VFA) were measured.

As a result, as shown in FIG. 7, it was confirmed that the VFA concentration increased as the culture progressed in the control and all the test zones.

Example  9. Depending on incubation time Shinny Green  Confirmation of content change

2, 4, 6, 8, 10, 12, and 24 hours) while inoculating the microorganism inoculum into the control and test sites according to Example 1-2 and culturing the cells. The cells were harvested by centrifugation (Brushless DC centrifugation (2,500 rpm x 15 min) using a centrifuge (VS-6000CF, Vision Scientific Co., Ltd., Korea). After centrifugation, the supernatant was taken and analyzed for the amount of cinnamin using HPLC.

As a result, as shown in Fig. 8, the peak of Sinnigreen was confirmed at 7.4 minutes. As a result of analyzing the content of sinigrin in 0 to 6 hours of culture, it was confirmed that the amount of sinigrin decreased with time as shown in Table 1 below.

[Table 1]

In addition, the supernatant was taken after centrifugation and the content of cinchinolytic hydrolyzate was analyzed by GC / MS.

As a result, as shown in the following Table 2, the hydrolysis products were not measured in all the test wells at 24 hours of incubation. Allyl isothiocyanate is a hydrolysis product of Sinnigreen, which is a volatile substance.

[Table 2]

Example  10. In animal models Sinny Green  Methane production Abatement  Check the effect

In order to confirm the effect of reducing the amount of methane produced by the shinigreen in vivo, the effect of reducing the methane production by the shinigreen was confirmed by reporting the black goat as an experimental animal.

More specifically, 4 black goats (5 times: 35.6 kg, 6 times: 39 kg, 7 times: 35.5 kg, 9 times: 39 kg) were weighed into a breathing chamber At which time the experiment proceeded in the respiratory chamber over the entire period, including the feed adaptation period. Prior to the experiment, timothy was fed free of diet. During the experimental period, 1.5% / day of each experimental animal body weight was divided into AM and PM twice. At this time, the four treatments including the control were carried out by adding 40 μ mol (T1), 60 μ mol (T2) and 120 μ mol (T3) of each Sinnigreen to the basic feed. The experimental period was set to 3 weeks, and the methane reduction effect on the treatments was measured for 3 days after the adaptation period.

As a result, as shown in Fig. 9, there was some data fluctuation during the three weeks of the adaptation period. However, during the three days of methane measurement, Sinni green 40 μ mol (T1), 60 mu mol (T2) (T3), and the average methane content of methane during the 3 days of methane measurement was about 1.18% and 60 μ mol, respectively, as compared with the control. T2) was about 8.95% and 120 mu mol (T3) was about 54.73%. As a result of the above experiment, the methane reduction effect was the largest in the 120 μ mol treatment, but 60 μ mol could be fed when the economic efficiency was taken into consideration.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (5)

A feed additive composition for reducing methane production in ruminants, comprising sinigrin as an active ingredient.
A feed composition for reducing methane production in a ruminant compounded with the feed additive composition of claim 1.
3. The method of claim 2,
Wherein the syngene is formulated at a concentration of 10 to 130 탆 ol.
A method of inhibiting methanogenesis in a ruminant, comprising feeding sinigrin to the animal.
5. The method of claim 4,
Characterized in that the syngene is fed at a concentration of 10 to 130 μmol.

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