JP2003088301A - Composition for suppressing methane production and feed composition for ruminants - Google Patents

Abstract

(57) [Summary] [PROBLEMS] Safe rumination without destroying the natural equilibrium of various microbial populations symbiotic with the digestive system of ruminants, and without remaining in host animals to reduce their commercial value Provided is a composition for suppressing methane production in an animal rumen. SOLUTION: By orally administering one or more selected from lactic acid bacteria and yeast derived from naturally fermented sheep's milk (laban) and oligosaccharides, methane production can be suppressed without deteriorating the rumen environment. . Further, by adding nitrate, the effect of suppressing methane production is enhanced. The lactic acid bacteria and yeasts are one or more microorganisms belonging to the genera Trichosporon, Candida, Leuconostoc, and Lactococcus. In particular, the oligosaccharide is preferably a galactooligosaccharide.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a composition for suppressing methane production for ruminants, more specifically, one or more kinds selected from lactic acid bacteria, yeasts and oligosaccharides and cysteine as an active ingredient. The present invention relates to a composition for suppressing methane production in the rumen of a ruminant.

[0002]

2. Description of the Related Art In ruminants such as cows and sheep, hydrogen produced during fermentation and digestion of structural carbohydrates is mainly used by methanogens symbiotic with the rumen to produce large amounts of methane. Will be a factor. The production of methane not only causes a loss of feed energy, but also has a considerable effect on global warming.

Examples of global warming substances include carbon dioxide, methane, chlorofluorocarbons, and nitrous oxide. Methane has a total of 0.
Although it constitutes only 4%, it has a heat absorption rate about 20 times that of carbon dioxide, and the effect is about 20% of total warming.
It contributes second only to carbon dioxide, and its amount is 1-yearly
It has increased at a rate of 1.3%, which is higher than carbon dioxide,
Reducing the amount of methane in the atmosphere is a global issue.

Particularly, the amount produced by digestive tract fermentation of livestock and wild animals is said to correspond to 16% of the total amount of methane emitted on the earth, and it is attracting attention as a source of methane. Also, in ruminants, the production of methane is 5-10% of the total calories obtained from the feed.
It is said to consume an amount equivalent to%. Usually, a plant as a feed is decomposed into hydrogen and carbon dioxide in the rumen by major fiber-decomposing bacteria, of which hydrogen is converted into methane by a methanogen and is discharged out of the body together with GEP. Converting this lost energy discharged outside the body into lower fatty acids or volatile fatty acids such as acetic acid, butyric acid, and propionic acid that ruminants use as nutrients leads to improvement in conversion efficiency of feed into meat. It is useful. In particular, acetic acid is said to cover about 70% of the total energy of ruminants, while propionic acid is considered to be an important carbon source for the production of tissues such as milk and meat. Therefore, if the amount of these organic acids can be increased, the feed efficiency is increased, which is economically preferable.

[0005] Conventionally, as a method for suppressing the production of methane in ruminants, a method of increasing concentrated feed such as cereals,
The method of administering a nitrate, a halogen compound, and a fatty acid was taken. However, the increase of the concentrated feed affects the protozoa (protozoa) inhabiting the rumen and causes a decrease in the amount of methanogens parasitic on the surface of the protozoa. Although the reduction in the amount of methanogenic bacteria reduces the rate of methane production, hydrogen remains, which not only reduces the activity of fiber-degrading bacteria, but also causes the frequent occurrence of metabolic diseases such as lactic acidosis. It is believed that. Furthermore, it is considered that an increase in acid concentration occurs and rumen parakeratosis (ruminal parakeratosis) develops.

[0006] Nitrate has a high affinity with hydrogen, and it is considered that by taking up hydrogen in competition with methanogens, the amount of hydrogen used for methane production is reduced and methane production is suppressed. However, there is a problem that a large amount of intake causes serious nitrite poisoning. Administration of halogen compounds and fatty acids also suppresses the production of methane, but depending on the intake amount and the intake period, problems such as chronic poisoning and reduction of fiber digestibility are caused. On the other hand, it has been reported that the use of ionophores such as salinomycin and monasin, which are antibiotics, suppresses the amount of methane produced, but there is concern that the emergence of resistant bacteria and their retention in livestock may occur.

[0007]

SUMMARY OF THE INVENTION Therefore, the present invention aims to reduce the commercial value of a microbial population that coexists in the digestive system of ruminants without destroying its natural equilibrium and remaining in the host animal. An object is to provide a safe composition for suppressing methane production in the rumen of a ruminant.

[0008]

Means for Solving the Problems As a result of intensive studies for solving the above problems, the present inventors have found that one or two selected from lactic acid bacteria and yeast derived from naturally fermented sheep milk (Laban) and oligosaccharides. It was found that the oral administration of more than one species can suppress the production of methane without deteriorating the lumen environment. It was also found that the addition of a nitrate further enhances the methane production suppressing effect.

[0009] In particular, it has been found that the combined use with nitrate not only enhances the effect of nitrate on suppressing methane production, but also suppresses the reduction of nitrate and suppresses the occurrence of nitrite poisoning. In addition, the effect of enhancing feed efficiency by suppressing methane production in the rumen of ruminants can be expected.

That is, the composition for suppressing methane production for ruminants according to claim 1 of the present invention is characterized by containing one or more kinds selected from lactic acid bacteria, yeast and oligosaccharides. The composition for suppressing methane production for ruminants according to claim 2, wherein the lactic acid bacterium and the yeast are one or more microorganisms belonging to the genus Trichosporon, the genus Candida, the genus Leuconostoc, the genus Lactococcus in the above-mentioned means. It is characterized by being.

The methane production inhibiting composition for ruminants according to claim 3 is characterized in that, in the means according to claim 1 or 2, the oligosaccharide is a galactooligosaccharide. The composition for suppressing methane production for ruminants according to claim 4 is selected from nitrate and / or cysteine, derivatives thereof and salts thereof (cysteines) according to any one of claims 1 to 3. It is characterized by containing one kind or two or more kinds. Furthermore, the feed composition for ruminants according to claim 5 is characterized in that the composition for suppressing methane production according to any one of claims 1 to 4 is added.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The lactic acid bacterium and yeast used in the present invention are not particularly limited as long as they have no pathogenicity.
Particularly preferred are microorganisms belonging to the genera Trichosporon, Candida, Leuconostoc and Lactococcus.

These lactic acid bacteria include Leuconostoc mesenteroides subspecies Mesenteroides L5 strain and Leuconostoc lactis L12.
Strain, Lactococcus lactis subspecies
Lactis L42 strain and the like. Examples of yeast include Candida kefir Y8A strain, Trichosporin sericium Y7A strain, and any lactic acid bacterium and yeast can be obtained from naturally fermented sheep milk (Laban) produced in Yemen. These L5 strain, L12 strain, L42
Strains, Y8A strains and Y7A strains are stored in the dairy chemistry laboratory, Department of Bioresource Chemistry, Faculty of Animal Science, Obihiro Storage University, in accordance with the provisions of Article 27-3 of the Patent Act.
It is sold to a third party as needed.

The dose of these microorganisms may be appropriately adjusted depending on the strain to be used and the animal to be administered.
~ 1.0 g / kg body weight, preferably 0.5-1.0 g / kg
Kg body weight may be administered. These microorganisms may be used alone or in combination of two or more. As the oligosaccharide used in the present invention, for example, galacto-oligosaccharide, raffinose, isolaffinose, lactulose, maltulose, trehalose, palatinose, maltooligosaccharide, isomaltooligosaccharide, soybean oligosaccharide, fructooligosaccharide, pectin oligosaccharide and the like can be preferably used.

The dosage of the oligosaccharide may be appropriately adjusted depending on the oligosaccharide used. For example, in the case of galactooligosaccharide, 0.5 to 2.0 g / kg with a purity of 80%.
Body weight, preferably 1.0 to 2.0 g / kg body weight may be administered. The oligosaccharides may be used alone or in combination of two or more.

When the oligosaccharide and the microorganism are used in combination, 0.05 to 1.0 g, preferably 0.5 to 1.0 g may be administered per 1 g of the microorganism. As the nitrate used in the present invention, potassium nitrate, sodium nitrate and the like can be used. These nitrates may be either chemically synthesized or extracted from natural products, and for example, nitrate-containing plants can be used as they are.

The dosage of nitrate is 0.05 to 1.0 g / kg ^0.75 (metabolized body weight), preferably 0.5 to 1.0 g / kg ^0.75 (metabolized body weight) if sodium nitrate is administered. Good.

As the cysteines or thiols used in the present invention, any of monomers, dimers, derivatives or salts thereof, or those in which a plurality of different thiols are bound are preferably used. However, cysteine is particularly preferable from the viewpoint of safety. The dose of cysteines or thiols may be appropriately adjusted according to the cysteines or thiols used, and the S equivalent is 0.0
05~0.21g / kg ^0.75 (metabolic body), preferably 0.05~0.21g / kg ^0.75 (metabolic body), it may be administered.

The timing of administration of these active ingredients is not particularly specified, and may be administered at any time as long as the feed remains in the lumen, but before administration of methane, the lumen may be administered. Since it is preferable that the active ingredient is present inside, it is preferable to administer the active ingredient immediately before or at the same time as feed administration. In particular, it can be efficiently administered by being mixed with feed.

These active ingredients may be administered individually or in the form of a mixture. The oligosaccharides, lactic acid bacteria, and bifidobacteria that are the materials of the present invention have been conventionally used in foods and the like, and have high safety, and therefore the dosage form can be arbitrarily selected, and added to and blended with feeds, pharmaceuticals, etc. be able to.

The material of the present invention may be used as it is, or may be mixed with a liquid or solid carrier, and if necessary, a solvent,
A dispersant, an emulsifier, a stabilizer, an excipient, a binder, a disintegrating agent, a lubricant, etc. are mixed, and for example, tablets, granules, powders, powders,
It can be made into a desired dosage form such as a capsule. Also,
Known feed additives such as bran may be added to the extent that the effects of the present invention are not impaired, and they may be used by mixing with livestock feed such as hay.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

[0023]

[Example 1] [Methane production inhibitory effect of lactic acid bacteria, yeast and oligosaccharide] (1) Preparation of bacterial cells For both lactic acid bacteria and yeast, strains derived from Yemen's naturally fermented sheep milk (Laban) were used. That is, as lactic acid bacteria, Leuconostoc mesenteroides subspecies
Mecenteroides L5 (hereinafter referred to as L5), Leuconostoc lactis L12 (hereinafter referred to as L12), Lactococcus lactis subspecies Lactis L23 (hereinafter referred to as L23), Lactococcus lactis subspecies Lactis L42 (hereinafter referred to as L42) was used. Examples of yeast include Candida kefir Y8A (hereinafter referred to as Y8A), Candida kefir Y4D (hereinafter referred to as Y4D), and Saccharomyces pastorianus Y3A.
(Hereinafter referred to as Y3A) and Trichosporin sericium Y7A (hereinafter referred to as Y7A) were used.

Each of the cells was cultured twice at 30 ° C. overnight twice, and the cells were cultured in a 100 cc medium for 2 nights and then spun down. The supernatant was removed and adjusted to 25 cc to obtain a sample cell.

(2) Adjustment of in vitro continuous methane production system An in vitro continuous methane production system was used as an inoculum (inoculum) from a lactating cow with rumen fistula according to the method of Takahashi et al. 750 ml of nylon cloth filtrate of collected rumen fluid and M
A mixture of 750 ml of cDougall artificial saliva was prepared as a medium.

As the substrate, 5 g of an orchard glass silage air-dried product was used. The test was carried out on 17 test groups of the oligosaccharide group, lactic acid bacterium group, yeast group, lactic acid bacterium + oligosaccharide group, yeast + oligosaccharide group, with the non-addition group as a control group.
As the oligosaccharide, GOS (β-1,4 galactooligosaccharide: manufactured by Yakult Honsha Co., Ltd.) was used, and 300 mg /
It was added at a rate of 1500 ml. After addition of the substrate and sample, the gas phase is collected at regular intervals and an infrared analyzer (Shimadzu V
IA-300, manufactured by Shimadzu Corporation) was used to measure the amount of methane produced according to a conventional method. The obtained results are shown in Tables 1 and 2.

[0027]

[Table 1]

[0028]

[Table 2]

As shown in Tables 1 and 2, as a result of comparing the total integrated amount of methane, Y7A was the most effective in the single addition zone of microorganisms, and then L5, Y8A, L42 and L12 showed the highest inhibitory effect in this order. It was On the other hand, Y4D, Y3A and L
In the 23 addition section, the methane production amount was higher than that in the control.

[0030]

[Example 2] [Effect of organic acid production by lactic acid bacterium, yeast and oligosaccharide] L5, L12 and L2 strains
3, L42, Y8A, Y4D, Y3A, Y7A,
GOS was used as the oligosaccharide. Sample bacterial cells were prepared in the same manner as in Example 1, and the same 17 test sections as in Example 1 were carried out using an in vitro continuous methane production system. After adding the substrate and the sample, the liquid phase was collected at regular intervals, and the organic acid concentration was measured by gas chromatography. That is,
Mixing metaphoric acid with the collected liquid and extracting / separating liquid 2
-Ethylbutyl butyric acid was quantified as an internal standard solution. The obtained results are shown in Tables 3 and 4.

[0031]

[Table 3]

[0032]

[Table 4]

As shown in Table 3, the addition of Y4D increased the total amount of organic acids. On the other hand, in L23, L42, and the yeast addition group, where the oligosaccharides were decomposed relatively quickly, the total amount of organic acids was increased by the combined use of oligosaccharides. Further, as shown in Table 4, the oligosaccharide alone reduced the proportion of acetic acid in the total organic acid, but the combined use of lactic acid bacteria and yeast Y8 increased the proportion of acetic acid. On the other hand, oligosaccharides, L5, L2
3, L49 and yeast and the combination of these strains with oligosaccharides increased the proportion of propionic acid.

[0034]

Example 3 [Methane production inhibitory effect of lactic acid bacteria, yeast and oligosaccharides in the presence of nitrate]
L5, L42 and Y8A are used, and GOS is used as the oligosaccharide.
Was used. In addition, sodium nitrate was used as the nitrate. The sample cells were prepared in the same manner as in Example 1, and the control group (no addition group), nitrate group, oligosaccharide group, L-cysteine group, nitrate + oligosaccharide group, nitrate + L-cysteine group,
Tests were carried out on 10 test groups of nitrate + lactic acid bacteria group and nitrate + yeast group using an in vitro continuous methane production system. In each test section, the gas phase was recovered over time, and the amount of methane produced was measured using an infrared analyzer as in Example 1. The results are shown in Table 5.

[0035]

[Table 5]

As shown in Table 5, methane production was suppressed in all test plots. The combined effect of nitrate and oligosaccharide was the highest, and then the combination of nitrate and L42,
Nitrate, a combination of nitrate and Y8A, a combination of nitrate and L5,
L-Cysteine, a combination of nitrate and L-cysteine, a combination of nitrate and Y7A, and an oligosaccharide showed a high inhibitory effect in this order.

[0037]

[Example 4] [Nitrite production inhibitory effect of L-cysteine, lactic acid bacteria, yeast and oligosaccharide] The test was carried out in the same manner as in Example 3. Using an in vitro continuous methane production system,
The diazo coupling method (Tak
ahashi, J.et.al, British Journal of Nutrition, 61:
741-748.1989.), And the amount of nitrite in each test section was measured. The results are shown in Table 6.

[0038]

[Table 6]

As shown in Table 6, the combined use of oligosaccharides, L-cysteine, Y8A and L42 suppresses the production of nitrite, and L-cysteine has the highest effect, followed by Y8A, oligosaccharides, The effect was higher in the order of L42. Also, L5 had no effect on nitrite production.

[0040]

Example 5 [L-cysteine in the presence of nitrate,
Effects of Lactic Acid Bacteria, Yeast, and Oligosaccharides on Organic Acid Production]
The test was conducted in the same manner as in Example 3. The amount of organic acid was measured in the same manner as in Example 2 from the liquid phase collected with time using an in vitro continuous methane production system. The obtained results are shown in Tables 7 and 8.

[0041]

[Table 7]

[0042]

[Table 8]

As shown in Table 7, nitrates, oligosaccharides,
And nitrates and oligosaccharides, L-cysteine, Y8A, L
The total amount of organic acids was increased by the combined use of 5 and L42. In particular, the effect of nitrate alone is high, followed by oligosaccharide, combination of nitrate and L-cysteine, combination of nitrate and Y8A,
The combined use of nitrate and L5, the combined use of nitrate and oligosaccharide, and the combined use of nitrate and L-42 showed high effects in this order. Table 8
As shown in, nitrates, and nitrates and oligosaccharides, L
-The combined use of cysteine, lactic acid bacteria and yeast increased the proportion of acetic acid in the total organic acids. On the other hand, the proportion of propionic acid increased when oligosaccharide alone, nitrate and Y7A, and nitrate and L42 were used in combination.

[0044]

[Example 6] [Preparation of feed] A feed having the following formulation was prepared, and GOS, L5, and Y8A, and nitrates and GO were prepared.
The combination with S, L5, and Y8A was added to make up 2% of the total feed weight. Barley 40 (% by weight) Wheat 30 Wheat feed 7.7 Peanut 6 Molasses 6.7 Salt 0.8 Limestone 0.4 Dipotassium phosphate 1.4 Barley vitamin 5 The obtained feed is highly feedable to livestock, Both health status and food intake were suitable.

[0045]

INDUSTRIAL APPLICABILITY According to the present invention, the production of methane in the rumen of ruminants can be suppressed and the production of useful organic acids such as acetic acid and propionic acid can be enhanced. Further, by using the nitrate and these materials in combination, it is possible to effectively suppress the methane generation while reducing the adverse effects of the nitrate.

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor So Chetla             Hokkaido, Obihiro-shi, Inada-cho, West Line 7, 7-30             Heights 209 (72) Inventor Budi Santoso             2-13 Inadacho Nishi, Obihiro City, Hokkaido International House             304 (72) Inventor Takeyoshi Arai             No. 15 livestock dormitory, West Inamachi West Line 2 Obihiro City Hokkaido             17 (72) Inventor Junichi Takahashi             2-6-13 Jiyugaoka, Obihiro City, Hokkaido (72) Inventor Kazumasa Kimura             1-1-19 Higashishimbashi, Minato-ku, Tokyo             Kurt Head Office (72) Harumi Mizukoshi, Inventor             1-1-19 Higashishimbashi, Minato-ku, Tokyo             Kurt Head Office F-term (reference) 2B005 BA08 JA04 KA04 LB07 MA08                 2B150 AA02 AB01 AC05 AC24 AD04                       DA49 DC15 DD15 DH13

Claims (5)

[Claims] 1. A composition for suppressing methane production for ruminants, which comprises one or more selected from lactic acid bacteria, yeast and oligosaccharides. 2. The lactic acid bacterium and the yeast are genus Trichosporon,
The composition for suppressing methane production for ruminants according to claim 1, which is one or more of microorganisms belonging to the genera Saccharomyces, Candida, Leuconostoc and Lactococcus. 3. The composition for suppressing methane production for ruminants according to claim 1, wherein the oligosaccharide is a galactooligosaccharide. 4. One or two selected from nitrates and / or cysteine, derivatives thereof and salts thereof (cysteines).
The composition for suppressing methane production according to any one of claims 1 to 3, which contains at least one species. 5. A composition for feed for ruminants, comprising the composition for suppressing methane production containing any one of claims 1 to 4.