JP2000325027A - Gain-promoter for animal - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a gain promoter for animals capable of maintaining normal enteric bacterial flora, rapidly increasing body weights and improving a feed efficiency ratio and disease prevention by containing a culture material of Clostridium buytyricum as an effective ingredient. SOLUTION: This gain promoter for animals contains a culture product or spores of Clostridium butyricum and an excipient such as corn starch. The Clostridium butyricum is preferably Clostridium butyricum NIP1020 strain (FERM BP-5794) or Clostridium butyricum NIP1021 strain (FERM BP-5795). The amount of the culture product or spores of the Clostridium butyricum to be added to the excepient is preferably 1-0.01 in weight ratio to 1 of the excipient when the culture product is used as an effective ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agent for promoting weight gain when given to livestock or the like to promote weight gain. More specifically, the present invention relates to butyric acid bacteria (Clostridi
um butyricum) or dried spores or kneaded and dried with appropriate excipients for livestock such as pigs and cattle, and poultry or cultured fish such as chickens. The present invention relates to an agent for promoting weight gain of livestock and the like, which promotes the body, contributes to normalization of the intestinal flora, and has an effect of preventing disease and improving feed efficiency.

[0002]

2. Description of the Related Art Clostridium butyricum is a spore-forming anaerobic Gram-positive bacillus that forms spores during the culturing process, and its mature spores are very stable. Butyric acid bacterium (Clostridium butyricum), even when given orally, does not die even in a highly acidic state such as gastric juice, reaches the intestinal fermentation site, proliferates in the intestinal tract, and is a lower volatile fatty acid mainly containing butyric acid. Produce antibacterial activity against pathogenic bacteria and the like, and contribute to maintaining normal intestinal flora.

In addition, 100 trillion, 100
Many and many types of microorganisms (fungi) inhabit, and constitute the intestinal flora while maintaining symbiotic or antagonistic relationship with each other,
It is involved in nutrient absorption and immunity, and is closely related to host health and disease. In general, if the diet and environment are constant and healthy, the intestinal flora balance of the animal is kept almost constant, but changes in nutritional components, stress, microbial infection, administration of antibiotics, etc. If the balance is lost due to the above factors, it may cause adverse effects on the health of the host.

[0004] On the other hand, in today's dairy industry in Japan, most of the feed ingredients for blending depend on imports from overseas, but soaring feed prices have put pressure on dairy farming. Therefore, for the purpose of reducing feed costs, there is a tendency that concentrated feeds using food wastes such as tofu lees, beer lees, and soy lees are frequently supplied. However, when the concentrated feed is fed in a large amount and the roughage is insufficient, for example, in cattle, abnormal fermentation occurs in the rumen, and due to the large amount of ammonia generated during the abnormal fermentation, the fermentation is resident in the stomach. However, there is a problem that the coliforms multiply in place of the microorganisms present, and the homeostasis of the microflora in the rumen is disrupted, causing nutritional disorders. In addition, pigs and chickens have been bred with multi-headed wings in order to improve productivity and labor saving under today's severe dairy management, and in such poor environment, the occurrence of infectious diseases and stress There is also a problem that diseases caused by hyperfunction or the like easily occur. In addition to the above-mentioned problems, drugs such as antibiotics used for disease control have problems such as persistence in meat and eggs and emergence of resistant bacteria, and their use tends to be regulated.

As described above, the ratio of feed costs to dairy management costs is very large, and it is necessary to increase feed efficiency in order to reduce the costs. In addition, various causes of livestock diseases are caused by normal intestinal diseases. Prevention can be achieved by maintaining the internal flora and exerting the biological defense function. Therefore, there is a strong need for a feed that can increase feed efficiency and maintain a normal intestinal flora.

[0006]

SUMMARY OF THE INVENTION Accordingly, the present invention relates to a feed additive for livestock and the like which can promote weight gain by increasing feed efficiency and prevent disease by maintaining a normal intestinal flora. It is an object of the present invention to provide a weight gain promoter.

[0007]

Means for Solving the Problems The present inventors have conducted intensive studies in order to solve these problems, and as a result, the produced substance shows an antibacterial action against pathogenic bacteria and the like and contributes to the maintenance of a normal intestinal flora. Butyric acid bacteria (Clostridi
um butyricum), butyric acid bacteria (Clostridium butyricum)
ricum), or by ingesting feed such as livestock animals with added spores recovered from the culture,
We found that the weight of the animals increased significantly, thereby completing the present invention.

[0008] That is, the above objects are as follows:
(E) is achieved.

(A) An animal weight gain promoter containing a culture of Clostridium butyricum as an active ingredient.

(A) An animal weight gain promoter comprising spores of Clostridium butyricum as an active ingredient.

(C) The above (A) further comprising an excipient.
Or the animal body weight-promoting agent according to (a).

(D) Clostridium butyricu
m) is Clostridium butyricum NIP1020 (Clo
stridium butyricum NIP 1020) (FERM BP-5)
794) The animal weight gain promoter according to any of (a) to (c) above, which is a strain.

(E) Clostridium butyricu
m) is Clostridium butyricum NIP1021 (Clo
stridium butyricum NIP 1021) strain (FERM BP-5)
795) The animal weight gain promoter according to any of (a) to (c) above, which is a strain.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The present invention relates to Clostridium butyricu
It is intended to provide an agent for promoting weight gain of livestock or the like containing the culture or spore of m) as an active ingredient.

The butyric acid bacterium (Clostridium b) used in the present invention is used.
utyricum) (hereinafter, parentheses are omitted) is not particularly limited as long as it belongs to Clostridium butyricum.
Butyricum NIP1020 (Clostridium butyricum N
IP 1020) strain (FERM BP-5794) and NIP1
021 (Clostridium butyricum NIP 1021) strain (FERM
BP-5795), and Clostridium butyricum ATCC 19398, ATCC 860, I
Although all Clostridium butyricum such as FO3315 and IAM 19001 strains can be used, of these, preferably, Clostridium butyricum N
IP1020 (Clostridium butyricum NIP 1020) strain (F
ERMBP-5794) and NIP1021 (Clostridi
um butyricum NIP 1021) strain (FERM BP-579)
5) is used.

In the present specification, the culture of butyric acid bacteria, which is an active ingredient of the weight gain promoter, includes both cultured cells of butyric acid bacteria and / or a culture solution.

The culture of butyric acid bacteria used in the present invention can be obtained by a known method, for example, the method disclosed in JP-A-08-252,088. Shows the embodiment in the following: the butyric acid bacteria, corn starch and amino acid solution each 2.0%, 10 ³ to 10 ⁷ calcium carbonate in the medium (previously autoclaved treatment) containing 0.75% / ml And incubate at 37 ° C. for 48 hours. If necessary, the culture solution thus obtained may be centrifuged (2,000 to 6,000 g × 10
(30 minutes) to separate the cultured cells of butyric acid bacteria. In addition, this cell is, for example,
Drying treatment by air drying or the like for 0 to 24 hours or 0 to 80 ° C,
The cells may be dried by subjecting them to a drying treatment under reduced pressure of 0.05 to 500 mmHg for 1 to 24 hours, for example, to form a dried product of the cells.

In the present invention, the medium used for culturing butyric acid bacteria varies depending on the kind of the strain used, etc., but a carbon source, a suitable amount of nitrogen source, inorganic salts, vitamins, etc. which can be used by the butyric acid bacteria used. As long as the medium contains other nutrients, either a synthetic medium or a natural medium may be used.

For example, the carbon source used in the medium according to the present invention is not particularly limited as long as the strain used can be assimilated. The carbon source is not necessarily limited to sugar, but in consideration of the growth of cells, a sugar or a sugar containing sugar that can be used by the bacteria to be used is preferably used. Specific examples of carbon sources that can be used include cellobiose, glucose, fructose,
Galactose, lactose, maltose, mannose,
Melibiose, raffinose, salicin, starch, sucrose, trehalose, xylose, dextrin, molasses and the like. Among these carbon sources, starch, glucose, fructose, sucrose and molasses are preferably used. The above carbon source,
One or more kinds may be selected and used in consideration of the butyric acid bacterium to be used. At this time, the addition concentration of the carbon source varies depending on the type of butyric acid bacterium and the carbon source to be used, the composition of the medium to be used other than the carbon source, and the like.
5 (w / v)%.

The nitrogen source and vitamins include:
For example, meat extract, peptone, yeast extract, soybean and wheat hydrolyzate such as taste liquid, soybean powder, milk casein,
Casamino acids, various amino acids, corn steep liquor,
Other examples include organic nitrogen compounds such as hydrolysates of animals, plants, and microorganisms, and ammonium salts such as ammonium sulfate. Among these nitrogen sources, peptone, yeast extract, meat extract, corn steep liquor and taste liquid are preferably used. The nitrogen source and vitamins described above,
In order to improve the growth of the butyric acid bacterium used, one or more kinds may be selected and used. At this time, the addition concentration of the nitrogen source varies depending on the type of the strain to be used, the type of the nitrogen source, and the medium composition other than the nitrogen source of the medium to be used, but when using peptone containing a large amount of the nitrogen source, it is usually , 0.5 to 4 (w / v)%, and when a taste liquid or corn steep liquor containing a large amount of nitrogen source and vitamins is used, usually 0.5 to 5 (w / v)%. Yes, and when a yeast extract or meat extract containing a large amount of vitamins is used, the content is usually 0.5 to 4 (w / v)%.

Further, as inorganic salts, magnesium,
One selected from phosphate, hydrochloride, sulfate, butyrate, propionate, acetate and the like of manganese, calcium, sodium, potassium, molybdenum, strontium, boron, copper, iron, tin and zinc, or Two or more can be used. In the medium, if necessary,
Antifoaming agents, vegetable oils, surfactants, drugs such as blood and blood components, vitamins and antibiotics, and physiologically active substances such as plant or animal hormones may be added as appropriate.

The culturing conditions used in the present invention vary depending on physiological properties such as the range of growth (pH, temperature, etc.) of the butyric acid bacterium used in the present invention. However, since butyric acid bacterium is obligately anaerobic, It is necessary that the cells be cultured under anaerobic conditions without aeration, with aeration of nitrogen or carbon dioxide gas, or by reducing the oxidation-reduction potential by adding a reducing agent to the medium. The culture conditions at that time are appropriately selected depending on the range of growth of the strain used, the composition of the culture medium, and the culture method, and are not particularly limited as long as the strain can grow. Specifically, the culture temperature is 20 to 42 ° C.

According to the present invention, the culture or spore of the above-mentioned butyric acid bacterium can be used as an active ingredient of a gain-promoting agent as it is, or can be dried and powdered as an active ingredient, or Kneaded with a suitable excipient and dried may be used as an active ingredient. Of these, the culture or spores of a butyric acid bacterium are dried and powdered, and the culture or butyric acid bacterium or The spores are kneaded with a suitable excipient, dried and powdered,
It is preferably used in the present invention. At this time, an example of drying and powdering a culture or spore of a butyric acid bacterium is shown below:
Reduce the pressure to 5 mmHg or less and freeze-dry for 5 to 72 hours.
At this time, the same treatment can be performed when spores are collected from the culture solution. Thus obtained to the active ingredient in, as butyric acid bacteria viable cultures dried product 1g per 10 ⁸ or more,
It contains 10 ⁹ or more per 1 g of recovered spores.

An example in which a culture or spore of a butyric acid bacterium is kneaded with an appropriate excipient and dried to be powdered is shown below: The culture solution obtained as described above is mixed with corn starch or the like. And the mixture is added at 7 mmHg
Reduce the pressure before and after and dry at about 50 ° C. for 3 to 10 hours. At this time, the same treatment can be performed when spores are collected from the culture solution. The active ingredient thus obtained contains 10 ⁵ or more butyric acid bacteria as viable bacteria per 1 g of dried culture, and 10 ⁸ or more per 1 g of recovered spores.

In the present invention, the excipient used for kneading and drying the culture or spore of the above butyric acid bacterium with an appropriate excipient is not particularly limited, and examples thereof include corn starch, starch, dextrin and the like. , Gluten, flour, bran, rice bran, soybean meal, glucose, lactose, fish meal, brewer's yeast, calcium carbonate, calcium silicate,
Examples include powdery substances generally used as feed materials, such as calcium phosphate, vermiculite, diatomaceous earth, and talc. Among them, corn starch, flour, calcium carbonate and diatomaceous earth are preferably used.

The amount of the culture or spore of butyric acid bacterium added to the excipient is usually 10 parts by weight relative to 1 excipient when the culture is used as an active ingredient.
To 0.001, preferably 1 to 0.01, and
When spores are used as the active ingredient, the weight ratio is usually 100 to 0.01, preferably 10 to 0.1, relative to the excipient 1.

The active ingredient used in the present invention can be administered as an agent for promoting body weight in livestock or the like as it is, or can be administered after it has been mixed with feed and can be administered more easily.
For example, it may be administered after processing into granules, tablets, pellets, flakes, blocks and the like. Of these, the processing step for processing into a dosage form that is easy to administer is performed using a known method by adding known excipients, binders, disintegrants, stabilizers, flavoring agents, and the like.

The feed to which the active ingredient according to the present invention is added is appropriately selected depending on the kind of animal to be administered, and specifically, cereals such as corn, wheat, milo; soybean, peanut, peas Potatoes such as potato, cassava, etc .; Vegetable oils such as soybean meal, rapeseed meal, flaxseed meal; Sugar bran such as rice bran, bran, starch meal, molasses meal, beer meal, tofu meal; fish meal And animal feeds such as skim milk powder, whey powder, feather meal and meat bone meal; and other feed ingredients that can be used as feed ingredients; and mixtures thereof. If necessary, vitamins, minerals, amino acids and the like may be added to the above-mentioned feed. At this time, the amount of the active ingredient used in the present invention to be added to the feed is not particularly limited as long as the above-mentioned effects can be obtained, but when the dried culture is used as the active ingredient, Is 0.0001% by weight or more, preferably 0.001% by weight.
1% by weight or more, and when the recovered spores are used as an active ingredient after drying,
It is at least 0001% by weight, preferably at least 0.0001% by weight. Further, when the culture is kneaded and dried with an appropriate excipient and used as an active ingredient, 0.1%
% By weight, preferably 1% by weight or more, and when the recovered spores are kneaded and dried with an appropriate excipient as an active ingredient, 0.0001% by weight or more, preferably 0.001% by weight or more.

The weight gain promoter of the present invention can be ingested without forcing the animal by forcing the subject animal to ingest the animal in a feed or the like. It contributes to the normalization of the intestinal flora and has various effects such as disease prevention and feed efficiency improvement. Therefore, the animals to which the body weight enhancer is administered are not particularly limited, and all animals in which such effects are desired can be administered. For example, domestic animals such as pigs, cows, sheep, and horses, chickens, quails, and ducks , Turkeys and other poultry, hamachi,
Cultured aquatic animals such as Thailand, flounder, coho salmon, puffer fish, rainbow trout, sweetfish, shrimp and the like can be mentioned.

[0031]

Next, the present invention will be described in more detail by way of examples.

Example 1 A medium (pH not adjusted) containing corn starch and an amino acid solution at 2.0% each and calcium carbonate at 0.75% was adjusted to 121%.
After autoclaving at 20 ° C. for 20 minutes, the medium was quenched with tap water.

In 900 ml of the above medium, Clostridium butyricum NIP1020 strain and NI
The P1021 strain was inoculated and cultured at 37 ° C. for 48 hours. Next, the resulting culture was freeze-dried to obtain a butyric acid bacterium culture as an active ingredient. Clostridium butyricum NIP thus obtained
The viable cell count of butyric acid bacteria in the active ingredient of the 1020 strain was measured, and found to be 7.1 × 10 ⁹ per 1 g of the dried product.

The spores were recovered by centrifuging the culture solution (6,000 × g, 30 minutes). 11.5 g of the spores thus recovered from the culture solution was added to corn starch 10.7 g as an excipient.
g, and the pressure was reduced to about 7 mmHg, followed by drying at room temperature for 24 hours to obtain a dried spore of butyric acid bacteria as an active ingredient. Clostridium butyricum NIP1020 strain and NIP102 thus obtained
The viable count of butyric acid bacteria in one active ingredient was measured, and found to be 1.4 × 10 ¹⁰ and 2.3 × 10 ¹⁰ per g of the dried product, respectively.

Example 2 As a test animal, a mouse (Std: ddY) (male, 5 weeks old)
Age: purchased from Japan SLC, Inc.). Ma
Instead, 20 mice were divided into 4 groups of 5 mice each. Then, each
The mice in the group were each exposed at a temperature of 20 ± 3 ° C. and a light of 12:00.
Bred in breeding cages under conditions that allow free access to food and tap water
Ingested. In this way, each of the prescribed foods is consumed
The day on which the mice started to be allowed to be 0 days, and the weight of the mice at this time (g)
Was measured. In addition, the feed was obtained from each of the effective samples obtained in Example 1.
Ingredients of butyric acid bacteria are 1 × 10 ⁸Individual / g dried product
Mixed with corn starch,
Solid feed for sale (MF, Oriental Yeast Co., Ltd.)
Used at about 0.05% by weight. Ma
In addition, without adding butyric acid bacteria culture or spore recovery,
Form feed (MF, Oriental Yeast Co., Ltd.) and water
A group to which only tap water was freely taken was set as a control group.

Mice were weighed (g) at weekly intervals after the start of ingestion. From this, the average weight in each group ±
The standard deviation (n = 5) was determined, and the results are shown in Table 1 below.

[0037]

[Table 1]

As is clear from Table 1, the group in which the culture of butyric acid bacteria or the recovered spores were added to the food showed a significant increase in the body weight of the mice (that is, the effect of gaining weight) as compared with the control group. And the difference in the gain effect was already observed at the first week of administration, and the difference became remarkable as the feeding period was extended. In addition, regarding the NIP 1020 strain, when the collected spores administration group and the culture administration group were compared, it was found that the culture administration group had a greater gain effect.

All the mice subjected to the test were healthy without any abnormality during the test period and two weeks after the test was completed. In addition, no abnormalities were found in the internal organs and the like in the autopsy findings two weeks after the end of the test.

[0040]

As described above, butyric acid bacteria (Clostridium b.
By giving the animal a body weight gain-promoting agent of the present invention containing, as an active ingredient, a culture of S. utyricum) or a spore recovered from the culture, a rapid increase in body weight (ie, a body weight gain effect) is observed. According to the present invention, butyric acid bacteria (Clo
It is well known that S. stridium butyricum grows in the intestinal tract, exhibits antibacterial action against pathogenic bacteria and the like and maintains a normal intestinal flora.
By ingesting the target animal with a culture-promoting agent containing a culture of ridium butyricum) or spores, effects such as normalization of intestinal flora, disease prevention, and improvement of feed efficiency can be expected.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (Reference) C12R 1: 145) (72) Inventor Itsuki Fujita 1240 Matsumaru, Isumi-machi, Isumi-gun, Chiba F-term within Niho Kagaku Co., Ltd. (Reference) 2B150 AA01 AA02 AA03 AA05 AB01 AB02 AB03 AB10 AC01 DD12 DD26 4B065 AA23X BC01 BD10 BD15 BD38 CA43

Claims (5)

[Claims] 1. An animal weight gain promoter comprising a culture of butyric acid bacteria (Clostridium butyricum) as an active ingredient. 2. An animal weight gain promoter comprising a spore of butyric acid bacterium (Clostridium butyricum) as an active ingredient. 3. The method according to claim 1, further comprising an excipient.
8. The animal weight gain promoter according to the above. 4. The method according to claim 4, wherein the Clostridium butyricum is Clostridium butyricum NIP1020 (Clostridium butyricum).
dium butyricum NIP 1020) strain (FERM BP-579)
4) The animal weight gain promoter according to any one of claims 1 to 3, which is a strain. 5. The method according to claim 5, wherein said Clostridium butyricum is Clostridium butyricum NIP1021 (Clostridium butyricum).
dium butyricum NIP 1021) strain (FERM BP-579)
5) The animal weight gain promoter according to any one of claims 1 to 3, which is a strain.