JP2002281912A - Feed composition suppressing generation of methane by ruminant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a methane generation inhibitor for ruminant free from problems of safety, handleability, or the like, and a feed composition for ruminant obtained by formulating the methane generation inhibitor. SOLUTION: The methane generation inhibitor for ruminant is characterized by comprising at least one kind of ingredient selected from eucalyptus oil, peppermint oil, Japanese horseradish oil, cineole, iodine and cyclodextrin clathrates of these materials as an active ingredient. The feed composition for ruminant is characterized in that the methane generation inhibitor is added thereto.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a feed composition for suppressing methane production in ruminants, and more particularly to eucalyptus oil, peppermint oil, wasabi oil, cineol, iodine and cyclodextrin clathrate compounds as active ingredients. The invention relates to a ruminant methane production inhibitor characterized by containing at least one substance selected from the group consisting of: and a ruminant feed composition to which the methane production inhibitor is added.

[0002]

2. Description of the Related Art In recent years, global warming has become a major problem. One of the causative substances is methane.
About 15% of the methane is produced from rumen of cattle, sheep and other ruminants. Research on the suppression of methane production from ruminants has been conducted for a long time, especially with ionophores (Goodrich, RD, J. Animal Sci., 1484-1498, 5
8, 1984) and halogen compounds (Chalupa, W., MPT Pres
s, England, 325-347, 1980), fatty acids (Blaxter, KL
et al., J. Sci. Fd. Agric., 17, 417-421, 1966) are known to have a very high ability to suppress methane production. However, ionophores are not only absorbed and affect host animals, but also have the potential to remain in milk, and have the disadvantage that halogen compounds and fatty acids are difficult to handle at room temperature, No effective control measures have yet been established.

Eucalyptus is an evergreen tree of the family Myrtaceae distributed in Australia and the like. Eucalyptus oil obtained by steam distillation from its leaves contains cineole as a main component and is known to have anti-inflammatory and antibacterial activities. Have been. Also, peppermint oil, wasabi oil,
Iodine is also known to have antibacterial activity. However, there has been no report on the use of such an antibacterial substance for suppressing methane production in ruminants. Further, eucalyptus oil, peppermint oil, and wasabi oil are volatile, so that they are difficult to handle at room temperature. Further, they are poorly water-soluble, so that their use in aqueous solutions is limited.

On the other hand, cyclodextrin (hereinafter referred to as CD)
May be abbreviated. ) Indicates that glucose is α-1,4
Α-, β-, and γ-CDs, which are cyclic dextrins linked by bonds and are composed of 6, 7, or 8 glucoses, are well known. Recently, a glucosyl group, a maltosyl group, a galactosyl group, a mannosyl group, a methyl group,
Modified CD with acetyl, hydroxypropyl, hydroxyethyl, monochlorotriazinyl, etc.
Have been synthesized. These CDs and modified CDs have a cavity inside the molecule, and since the inside of the cavity is hydrophobic, it has an inclusion function and has a property of taking in various oily substances. Therefore, utilizing this property, (1) stabilization of unstable substances, (2) retention of volatile substances,
Various uses such as (3) masking of off-flavors and (4) solubilization of difficult and insoluble substances are considered, and are used in a wide range of fields such as foods, pharmaceuticals, cosmetics, and industrial products.

[0005]

DISCLOSURE OF THE INVENTION As described above, an object of the present invention is to consider safety and handling properties in view of the fact that a substance effective for suppressing methane production by ruminants has not been established. An object of the present invention is to provide a ruminant feed composition for ruminants, which contains no problem in terms of methane production inhibitor for ruminants and the methane production inhibitor.

[0006]

Means for Solving the Problems The present inventors have focused on various antibacterial substances and the inclusion effect of CD described above, and have been developing various feed compositions capable of suppressing methane production by ruminants. As a result of repeated studies, antibacterial substances such as eucalyptus oil, peppermint oil, wasabi oil, cineol, and iodine are useful. In addition, these substances are included in a CD and powdered into a stable clathrate. That the powder is obtained, that the powder has an effect of improving poor water solubility, and that adding this clathrate powder to a mixed microorganism sample collected from bovine rumen suppresses methane production, The present invention has been completed based on these findings.

That is, the present invention is characterized in that it contains at least one substance selected from eucalyptus oil, peppermint oil, wasabi oil, cineol, iodine and cyclodextrin inclusion compounds as active ingredients. The present invention relates to a ruminant methane production inhibitor and a ruminant feed composition comprising the ruminant methane production inhibitor.

[0008]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Eucalyptus oil, peppermint oil and wasabi oil used in the present invention are all mixed oils, while eucalyptus oil is mainly composed of cineole and peppermint oil is mainly composed of menthol. As for the wasabi oil, those containing allyl isothiocyanate as a main component are preferably used. The antibacterial substances to which cineol and iodine are added may be used alone or in combination of two or more. Furthermore, these CD inclusions can be used as well. Since the inclusion and the powdering with CD have various characteristics as described above, a better effect can be expected as compared with the case where the antibacterial substance itself is used.

The CD used in the present invention is α-CD,
In addition to unmodified CDs such as β-CD and γ-CD, these may be glucosyl, maltosyl, galactosyl, mannosyl, methyl, acetyl, hydroxypropyl, hydroxyethyl, monochlorotriazinyl, etc. Any of the modified CDs obtained by the modification described above, or a mixture thereof may be used.

The method for producing the CD inclusion compound of the above antibacterial substance which can be used in the present invention is exemplified as follows.
5-100% (w / v), preferably 1-50% CD
To the aqueous solution, 0.01 to 10 times, preferably 0.05 to 5 times the amount of the guest is added to the CD, and a solution obtained by mixing with a stirrer such as a homogenizer is used in a commonly used freeze dryer or spray drying. It can be obtained by pulverizing with a machine.

The methane production inhibitor for ruminants according to the present invention can be used in various forms, for example, powder,
Various forms such as granules and tablets can be mentioned, and an excipient, a bulking agent and the like can be added as needed. The amount of the active ingredient in the methane production inhibitor may be determined in consideration of the purpose of use and the like, but is usually 0.1 to 80.
% By weight, preferably 1 to 20% by weight.

Next, the feed composition for ruminants according to the present invention is a feed composition to which the above-mentioned methane production inhibitor is added, and the mixing ratio of the methane production inhibitor in the feed composition is usually 0.1 to 10% by weight, preferably 1 to 5
% By weight. The feed composition for ruminants is not particularly limited, and commercially available products may be used as they are, or if necessary, silage, hay or the like may be added to the commercially available products. Illustrative examples of feed compositions for cattle include corn silage 40%, Sudan grass hay or alfalfa hay cubes 14%,
Some commercial feeds consist of 46%. As an example of a commercially available compound feed, a feed having a crude protein of 16% or more and a digestible nutrient amount of 71% or more (trade name: dairy cow flake 1)
6, Nissin Feed Co., Ltd.). on the other hand,
Illustrating the feed composition for sheep, Timothy hay
Some consist of 55%, 24% alfalfa hay cubes, and 20% commercial formula feed. One of the commercial formula feeds
For example, 11% crude protein, total digestible nutrients
74% of feed (trade name: beef cattle fattening, all dairy chain). In the present invention, the feed composition containing the methane production inhibitor may be freely taken by ruminants. In addition, it can be taken for a long period of time.

[0013]

EXAMPLES Next, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples. Test Example 1 (1) Preparation of Inclusion Compound of Methane Production Inhibitor by α-CD Eucalyptus oil 50 m in 2000 ml of 10% α-CD aqueous solution
L was added, and the mixture was stirred at room temperature with a homogenizer, and the obtained precipitate was dried by vacuum drying or spray drying to obtain a eucalyptus-α-CD clathrate. (2) Measurement of component content of clathrate compound Eucalyptus-α-CD clathrate compound 5 produced in (1) above
0 mg was precisely weighed, placed in a separating funnel, and dissolved in 140 mL of water. Furthermore, 10 mg of carvone is used as an internal standard.
And 20 mL of ether were added, and the mixture was shaken vigorously and allowed to stand. The upper ether layer is subjected to gas chromatography (GC-14A,
(Manufactured by Shimadzu Corporation). That is, using a capillary column (TC-1 60 m × 0.25 mm), the column temperature was set at 100 ° C. and the detector temperature was set at 300 ° C. Using helium gas as a carrier gas, the components were measured by a flame ionization detector. The peak area of each component was automatically measured by a chromatopack (manufactured by Shimadzu Corporation). The content of each component was calculated from a separately prepared calibration curve. As a result, the yield of the eucalyptus-α-CD inclusion compound was 170
g, yield 68%, eucalyptus content about 7%.

Example 1 (1) Sampling of Samples A catheter was orally inserted into the lumen of a dairy cow fed with roughage and concentrated feed, and about 1 L of the rumen contents was sucked and collected in a filter bottle. Immediately after the collection, the rumen contents were stirred and filtered with a double gauze, and the filtrate was used as a microorganism-mixed sample. (2) Culture method The microorganism mixture sample obtained in the above (1) was doubled in a salt solution (g / L: KH ₂ PO ₄ 3; Na ₂ HPO ₄ .12H ₂ O 1).
8; NaCl 1; NaHCO ₃ 1; MgSO ₄ .7H ₂
O 0.2; after dilution with CaCl ₂ .2H ₂ O 0.2), 60
mL was placed in a culture tube. Furthermore, before culture, 0.6 g / L of cysteine hydrochloride and Eucalyptus α-CD were used as reducing agents.
Inclusion compounds and carbohydrates (glucose, cellobiose,
(0.12 g / vial) was added thereto, followed by shaking culture at 38 ° C. for 6 hours under anaerobic conditions. The eucalyptus-α-CD inclusion compound was added in an amount of 5 to 40 mg as eucalyptus oil.

(3) Measurement of methane production A gas chromatograph (GAS CHROMATOGRAP) with a thermal conductivity detector
H GC-8A (manufactured by Shimadzu Corporation) and a column for measuring methane (Molecular Sieve 5A 60-80 mesh, 1.6 mx 3.2 m)
m), the column temperature was set to 60 ° C. and the detector temperature was set to 80 ° C., and argon was used as a carrier gas. The peak area of methane was automatically calculated by the aforementioned chromatopack. The content of each component was calculated from a separately prepared calibration curve. (4) Measurement of Volatile Fatty Acid (VFA) Concentration 2 mL of the supernatant of the culture solution was taken into a test tube, and a 3N-H ₂ SO ₄ solution containing 12% metaphosphoric acid as a deproteinizing agent was 0.4 m.
L was added, covered with a rubber stopper, and stored in a refrigerator. After standing overnight, the mixture was centrifuged for 10 minutes in a cooling centrifuge (Model M-160-SAKUMA) set at a temperature of 4 ° C. and a rotation speed of 5000 rpm, and the supernatant was used as a sample for analysis. 1 μL of the supernatant is applied to a gas chromatograph (GAS CHROMA) using a flame ionization detector.
Injected into TOGRAPHGC-8A, manufactured by Shimadzu Corporation, and used for VFA measurement column (FAL-M SHINCARBON A 80-100 mesh, 2.1mx
3.2mm), column temperature 130 ° C, detector temperature 2
The temperature was set to 00 ° C., and helium was used as a carrier gas. The peak area of each gas was automatically calculated by the aforementioned chromatopack. The content of each component was calculated from a separately prepared calibration curve.

(5) Measurement of the number of protozoa The number of protozoa of the fixed and stained sample was measured using a hook rosenthal computer. Using an optical microscope (Nikon Type 104), it was visually measured at a magnification of 10 × 10 and converted to the number in 1 mL of the rumen solution. This method followed the “Clinical Pathological Examination Guidelines for Livestock Mutual Aid” (edited by the Ministry of Agriculture, Forestry and Fisheries Economic Bureau, National Agricultural Mutual Aid Association, 1997). (6) The obtained results are shown in FIGS. That is, FIG. 1 shows the amount of methane produced after culturing for 6 hours, FIG. 2 shows the number of protozoa, FIG. 3 shows the amount of acetic acid produced, and FIG. 4 shows the amount of propionic acid produced. In addition, these are relative values when the case where no methane production inhibitor is added is set to 100. As is clear from the results shown in FIGS. 1 to 4, when 5 to 40 mg of eucalyptus / α-CD clathrate was added as a eucalyptus oil to the mixed microorganism sample, methane generation and the number of protozoa decreased as the amount of addition increased. did. Acetic acid and propionic acid also increased. From these results, Eucalyptus-α-CD inclusion compound reduces methane production by suppressing protozoa activity,
It was also shown to improve feed efficiency.

Example 2 The procedure of Example 1 was repeated except that 5 to 15 mg of peppermint oil was added as the peppermint oil. The obtained results are shown in FIGS. That is, FIG. 5 shows the amount of methane produced after culturing for 6 hours, and FIG. 6 shows the number of protozoa. In addition, these are relative values when the case where no methane production inhibitor is added is set to 100. As is apparent from these results, the methane generation and the number of protozoa decreased as the amount of addition increased. Thus, the peppermint-α-CD inclusion compound reduces methane production by suppressing protozoa activity,
It has been shown to improve feed efficiency.

Example 3 The same procedure as in Example 1 was carried out except that 5 to 15 mg of peppermint / β-CD inclusion compound was added as peppermint oil. The obtained results are shown in FIGS. That is, FIG. 7 shows the amount of methane produced after culturing for 6 hours, and FIG. 8 shows the number of protozoa. In addition, these are relative values when the case where no methane production inhibitor is added is set to 100. As is apparent from these results, the methane generation and the number of protozoa decreased as the amount of addition increased. Therefore, the peppermint-β-CD inclusion compound also reduces methane production by suppressing protozoa activity,
It has been shown to improve feed efficiency.

Example 4 1-10 mg of wasabi / CD clathrate as wasabi oil
Except having added, it carried out similarly to Example 1. In this example, the CD is "K50" (a CD mixture of 50% CD and 50% dextrin;
-CD containing 30% or more). The obtained results are shown in FIGS. That is, FIG. 9 shows the amount of methane produced after culturing for 6 hours, and FIG. 10 shows the number of protozoa. As is apparent from these results, the methane generation and the number of protozoa decreased as the amount of the wasabi / CD inclusion compound increased. Therefore, it was shown that the wasabi-CD inclusion compound also reduced the amount of methane produced by suppressing protozoa activity and improved feed efficiency.

Example 5 Cineol / α-CD clathrate was used as cineol.
The same procedure as in Example 1 was performed except that 40 mg was added. The results are shown in FIGS. FIG. 11 shows the amount of methane produced after culturing for 6 hours, and FIG. 12 shows the number of protozoa. As is evident from these results, the methanation and the number of protozoa decreased as the added amount increased. Accordingly, it was shown that the cineol-α-CD inclusion compound also reduced the amount of methane produced by suppressing protozoa activity and improved feed efficiency.

Example 6 1-5 mg of iodine / β-CD clathrate as iodine
Except having added, it carried out similarly to Example 1. The results are shown in FIGS. FIG. 13 shows the amount of methane produced after culturing for 6 hours, and FIG. 14 shows the number of protozoa. As is evident from these results, the methanation and the number of protozoa decreased as the added amount increased. Therefore, it was shown that the iodine / β-CD inclusion compound reduces the amount of methane produced by suppressing protozoa activity and improves feed efficiency.

[0022]

Industrial Applicability The methane production inhibitor for ruminants of the present invention is superior in safety to conventionally known ionophores and the like since the active ingredient is a natural product or an extract thereof. In addition, by enclosing these substances with CD, not only long-term storage can be performed, but also advantages such as solubilization of the active ingredient and masking of off-flavors can be obtained.

Therefore, by ingesting a ruminant feed composition containing the methane production inhibitor into ruminants, it is possible to effectively suppress methane production in the rumen of the ruminant and improve feed efficiency. Can be.

[Brief description of the drawings]

FIG. 1 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of methane production in Example 1.

FIG. 2 is a graph showing the relationship between the amount of a methane production inhibitor added and the number of protozoa in Example 1.

FIG. 3 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of acetic acid produced in Example 1.

FIG. 4 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of propionic acid produced in Example 1.

FIG. 5 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of methane production in Example 2.

FIG. 6 is a graph showing the relationship between the amount of a methane production inhibitor added and the number of protozoa in Example 2.

FIG. 7 is a graph showing the relationship between the addition amount of a methane generation inhibitor and the amount of methane generation in Example 3.

FIG. 8 is a graph showing the relationship between the amount of a methane production inhibitor added and the number of protozoa in Example 3.

FIG. 9 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of methane production in Example 4.

FIG. 10 is a graph showing the relationship between the added amount of a methane production inhibitor and the number of protozoa in Example 4.

FIG. 11 is a graph showing the relationship between the amount of a methane production inhibitor added and the amount of methane produced in Example 5.

FIG. 12 is a graph showing the relationship between the amount of a methane production inhibitor added and the number of protozoa in Example 5.

FIG. 13 is a graph showing the relationship between the amount of the methane generation inhibitor added and the amount of methane generation in Example 6.

FIG. 14 is a graph showing the relationship between the amount of a methane production inhibitor added and the number of protozoa in Example 6.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Noriko Ajizaka 13-46 Ogurocho, Tsurumi-ku, Yokohama, Kanagawa Prefecture Inside the Yokohama International Biotechnology Research Institute Co., Ltd. (72) Koji Hara 13-46 Ogurocho, Tsurumi-ku, Yokohama-shi, Kanagawa Inside Yokohama International Bio Research Institute Co., Ltd. (72) Katsuhiko Mikuni 13-46 Ogurocho, Tsurumi-ku, Yokohama City, Kanagawa Prefecture Inside Yokohama International Bio Research Institute Co., Ltd. (72) Inventor Hisao Itabashi 1-39-1 F, Tobita Supply, Chofu City, Tokyo Terms (reference) 2B005 BA01 BA05 BA06 BA07 2B150 AA02 AB20 DD31 DH17 DJ01

Claims (2)

[Claims] 1. A ruminant animal comprising as an active ingredient at least one substance selected from eucalyptus oil, peppermint oil, wasabi oil, cineol, iodine and cyclodextrin inclusion compounds. Methane production inhibitor. 2. A ruminant feed composition comprising the ruminant methane production inhibitor according to claim 1 added thereto.