JPH06181696A - Method for promoting growth of animal and powdery pharmaceutical preparation of dead microbial cell microorganism belonging to genus clostridium - Google Patents

Abstract

PURPOSE:To safely and effectively promote the growth of an animal such as livestock, fowl or a cultured aquatic animal without a fear of residual medicines, etc., and side effects by orally administering a dead microbial cell of a microorganism belonging to the genus Clostridium to an animal. CONSTITUTION:A microorganism belonging to the genus Clostridium [e.g. Clostridium.butyricum.MIYAIRI (FERM P-2789)] is inoculated into a culture medium, cultured at 30-40 deg.C for 14-24hr and then sterilized by heat treatment or treatment with formalin or other germicides to provide a nutrient dead microbial cell. The nutrient dead microbial cell of the Clostridium.butyricum.MIYAIRI in an amount of 0.00005-80wt./wt.% is subsequently blended with 0.001-50wt./wt.% saccharides, 0.015-90wt./wt.% starch, 0.0005-33wt./wt.% sparingly water-soluble inorganic substance and 1-99.9wt./wt.% excipient to prepare a pharmaceutical preparation having >=10<6> cell number of the nutrient dead microbial cells based on 1g pharmaceutical preparation. The obtained pharmaceutical preparation is then orally administered to an animal such as livestock, fowl or a cultured aquatic animal to effectively promote the growth of the animal.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for promoting animal growth and a powdered preparation of killed Clostridium spp.

[0002]

2. Description of the Related Art Livestock,
In poultry and aquaculture aquatic animals, antibacterial substances such as antibiotics have been widely added to feeds for the purpose of promoting growth.
However, recently, it has been frequently pointed out by consumers and mass media that antibacterial substances, etc. remain in livestock and cultured fish, and the use of antibacterial substances is being reviewed from the viewpoint of safe food supply to the people. is there. On the other hand, probiotics have been used for a long time for the purpose of maintaining the health of mammals such as humans and livestock. It is said that the action of this probiotic agent is due to an antagonistic action against harmful bacteria and the like. Therefore, it is considered that living bacteria are essential as the probiotic agent.

In the case of a probiotic agent, the temperature cannot be raised due to the problem of protein denaturation, and strong crumples and pellets cannot be produced. In a dry feed such as eel feed, the enzyme produced by viable bacteria causes the feed to deteriorate. Further, when the viable bacterium is a spore-forming bacterium, a treatment for forming a spore is required in order to prevent the generation of dead bacterium as much as possible, so that the yield is poor. Therefore, an object of the present invention is to provide a safe growth-promoting substance that replaces the antibacterial substance and the probiotic agent.

[0004]

DISCLOSURE OF THE INVENTION The present invention provides a method for promoting growth of an animal, which comprises orally administering killed cells of a microorganism belonging to the genus Clostridium to an animal, and a vegetative cell killed with microorganisms belonging to the genus Clostridium. The present invention relates to a powdered preparation of killed Clostridium spp. Cells containing the body and sugars and / or starch as essential components.

Examples of microorganisms belonging to the genus Clostridium include Clostridium butyricum ( Clost
ridium butyricum ). Particularly preferred are, for example, Mycobacterium miyali. The bacterium Miyari is scientific name Clostridium butyricum
Clostridium butyricu
m MIYAIRI), which is currently marketed as a probiotic agent by Miyarisan Co., Ltd. and has no side effects even when administered to humans or livestock for many years. As one kind of this Mycobacterium, there are Clostridium butyricum Miyairi 588 strain (Kikuko Kenjo No. 2789: FERM BP-2789). There is no limitation on the viable bacteria for producing dead cells, whether in the spore formation stage or in the vegetative stage, but the vegetative stage is preferable. Production method of live bacteria is USP 489273
1 is disclosed.

The dead cells used in the present invention can be obtained by a conventional method such as heat treatment, formalin treatment or other treatment method using a bactericide or the like. Further, it may be substantially dead cells, and live cells may be used as long as it is substantially dead cells during formulation. To produce the trophozoites of microorganisms belonging to the genus Clostridium, the microorganisms belonging to the genus Clostridium are inoculated into a suitable medium such as glucose-containing ordinary broth without any particular limitation on the method of culturing the cells and the method of separation, and the culturing is carried out at 30 ° C or higher. Preculture at 40 ° C. for 14 hours to 24 hours. The liquid after this culturing is inoculated into the same medium as the scale-up pre-culture, and the main culture is performed at the same temperature and time. After completion of the main culture, the cells may be collected using an appropriate device such as a centrifuge. The medium is not particularly limited as long as it is for the bacterium, and for example, ordinary broth for Clostridium butyricum bacterium (meat extract 0.8%,
Although any medium such as peptone 1% and sodium chloride 0.5%) can be used, a liquid medium containing glucose, yeast extract and peptone is preferable. In addition, Japanese Examined Sho 37-8
In the method disclosed in No. 300, the culturing time may be shortened (for example, about 14 to 24 hours), the culturing may be suspended in the state of the vegetative body before spore formation, and the bacterial cells may be collected.

In order to obtain the trophozoite powder of the microorganism belonging to the genus Clostridium used in the present invention, the cultured vegetative cells are filtered in the air, or when it is not enough. Furthermore, it is sufficient to suspend the filtrate in water containing oxygen (Clostridium bacteria are obligate anaerobes, so their vegetative body is killed only by touching the air to become vegetative dead cells). . The trophozoite killed cells can be obtained by a normal method of live vegetative cells, for example, a heat treatment, a formalin treatment, or a treatment method using a bactericide or the like. Anything that can be said to be substantially dead cells,
Live cells may be used as long as they are substantially dead cells during formulation.

The dead cells used in the present invention include dead cells 1
The number of bacterial cells per g is 10 ⁹ or more, preferably 10 ⁹ to 10 ^11.
It only needs to be included. The bacterial cell number is a value obtained by counting a cell suspension containing a bacterial cell emulsified in physiological saline in a hemocytometer with a microscope. In the preparation of the present invention, 10 ⁶ / g or more, preferably 10 ⁶ to 10 ¹¹ / g
g, and more preferably about 10 ⁶ to 10 ¹⁰ cells / g of dead cells sterilized with live cells.

The animals to be administered are non-human animals,
Examples include livestock, poultry, and aquaculture aquatic animals. As the preparation, the bacterial cells are sterilized after culturing, and only the dead bacterial cells are separated and concentrated, and the dried dead bacterial cell bulk powder can be directly used as a preparation, or rice bran oil cake, flour, glucose, silicic acid anhydride, The formulation can be prepared by mixing with any excipients approved by the feed safety law such as bran. Further, a culture broth cultivated as an active ingredient, including the residue, is concentrated and dried, and a sterilized cell-containing material can also be used. Examples of the form of the preparation include powders, tablets, pellets, fine granules, capsules and the like. When a tablet is produced, it may be prepared by a conventional method using an excipient, a disintegrating agent, a binder, a lubricant, etc., if necessary.

Examples of the excipient include sugars such as lactose, D-mannitol, D-sorbitol and sucrose, starches such as corn starch and potato starch, inorganic salts such as calcium phosphate, calcium sulfate and precipitated calcium carbonate. can give.

Examples of the disintegrant include hydroxypropyl starch, sodium carboxymethyl starch,
Examples thereof include starches such as partially pregelatinized starch, cellulose derivatives such as carboxymethylcellulose calcium, carboxymethylcellulose and low-substituted hydroxypropylcellulose, and other synthetic polymers having polyvinylpyrrolidone in a crosslinked structure.

Examples of the binder include polymers such as polyvinylpyrrolidone, hydroxypropylcellulose, hydroxypropylmethylcellulose, gelatin and gum arabic. Examples of the lubricant include talc, waxes, natural products such as light anhydrous silicic acid and derivatives thereof, stearic acid, magnesium stearate, calcium stearate, aluminum stearate, sucrose fatty acid ester sugar fatty acids and the like. Examples thereof include metal salts. In addition, other high molecular compounds such as macrogol are appropriately used for tablets. When the preparation of the present invention is used for promoting the growth of animals, it is usually added to feed or drinking water, and therefore, it is preferable to use a powder preparation. In this case, the dead cell powder of the microorganism belonging to the genus Clostridium in the preparation is 0.00005 to 80%, preferably 0.0005 to 70%, more preferably 0.01 to 50%.
% And the balance is carrier.

In the preparation of the present invention, a combination of vegetatively killed bacterial cells with sugar and / or starch is preferred because the growth promoting action is further enhanced. That is, this preparation is a powdered Clostridium cell death preparation containing, as essential components, vegetative cells killed by microorganisms belonging to the genus Clostridium and saccharides and / or starch. The sugar used in the powdered preparation of killed cells may be any sugar such as glucose, fructose, sucrose, oligosaccharide, lactose, mannitol, sorbitol, xylitol, etc., but lactose is preferable. Also, starch is wheat starch,
Any potato starch, corn starch or other starch may be used, but corn starch is preferred. It is sufficient that the vegetative dead bacterial powder of a microorganism belonging to the genus Clostridium contains the number of cells of 10 ⁹ or more, preferably about 10 ⁹ to 10 ¹¹ / g.

In order to produce the Clostridium spp. Killed cell powder preparation of the present invention, for example, sugar or starch powder may be added to the killed cell powder and simply mixed, or the wet killed cells may be mixed with sugar or starch. Although starch may be added and dried and ground, in view of easiness of operation, after adding sugar and / or starch to dead cells and suspending them in water, the inlet temperature of hot air is 120 to 200 ° C., preferably 130. It is better to carry out spray drying at ˜170 ° C., outlet temperature 30˜150 ° C., preferably 50˜100 ° C. The amount of sugar or starch added is 0.04 to 700 parts by weight of sugar with respect to 1 part by weight of dead microbial cells of a microorganism belonging to the genus Clostridium.
Preferably 0.1 to 70 parts by weight, starch is 0.2 to 700
Parts by weight, preferably about 0.4 to 70 parts by weight. In the case of spray drying, the operation becomes easy by adding a poorly water-soluble physiologically acceptable inorganic substance such as calcium carbonate. In this case, the amount of the inorganic substance added is 0.005 to 0.5 part, preferably 0.01 to 0.5, based on the total amount of the above essential components.
0.2 parts, and more preferably 0.01 to 0.1 parts.

Clostridium obtained by the above method
The ratio of each component of the powdered formulation of killed genus bacteria is
0.05-80% of dead vegetative cells of microorganisms belonging to the genus Mumu,
Preferably 0.5-70%, more preferably 5-50
%, Sugar 1-50%, preferably 3-40%, more preferably
5 to 30%, starch 15 to 90%, preferably 25
-80%, more preferably about 35-75%,
10 g of dead microbial cells per 1 g of the preparation⁹More than one, preferred
Kuha 10⁹-10¹¹More preferably 10 ⁹-10
^TenIt only needs to be included. When using inorganic materials
In that case, its proportion in the formulation is 0.5 to 50%, preferably 1
-20%, more preferably 1-10%.

An excipient powder may be further added to the above powder formulation. Examples of the excipient include defatted rice bran, soybean flour, bran, chaff, calcium carbonate, sugar, starch, brewer's yeast, wheat flour, and any other excipient approved by the Feed Safety Law. Not only one of these excipients, but two
You may use together 1 or more types. The addition ratio is 0.01 to 1000 parts by weight, preferably 0.1 to 800 parts by weight, and more preferably 1 to 500 parts by weight, per 1 part by weight of the above powder preparation. The amount of dead cells in this excipient-containing powder formulation is
The number of bacterial cells per gram of the preparation is 10 ⁶ or more, preferably 10 ⁶ to
The number is about 10 ¹¹ , more preferably about 10 ⁶ to 10 ¹⁰ .

The proportion of each component in the powdered preparation containing excipients will be described. 0.00005-80%, preferably 0.0005-5%, of dead microbial cells of microorganisms belonging to the genus Clostridium.
70%, more preferably 0.01 to 25%, sugar 0.0
01 to 50%, preferably 0.003 to 40%, more preferably 0.01 to 15%, starch 0.015 to 90
%, Preferably 0.03-80%, more preferably 0.05-40%, excipient 1-99.9%, preferably 10-99.8%, more preferably 50-99.7.
%, The degree. When a spray-dried preparation containing an inorganic substance is used as the powder formulation, the proportion of the inorganic substance is 0.0005 to 33%, preferably 0.001 to 15%.
%, And more preferably about 0.002 to 5%.

When sugar, starch, or an inorganic substance is used as the excipient, the ratio of each component in the preparation is 0.001 to 99.9 as the total amount of the components in the excipient and the dry powder.
%, Preferably 0.003 to 99.8%, more preferably 0.01 to 99.7%, starch 0.015 to 99.9.
%, Preferably 0.03 to 99.8%, more preferably 0.05 to 99.7%, and inorganic substances 0.0005 to 99.%.
It is about 9%, preferably 0.001 to 99.8%, and more preferably about 0.002 to 99.7%.

The formulation of the present invention may further contain auxiliaries such as a fluidity improver, anticaking agent and anti-scattering agent. The preparation of the present invention may be directly administered to animals such as humans and livestock, but is usually added to animal feed such as livestock, poultry or aquaculture aquatic animals, or suspended in drinking water for administration. Administration is as long as necessary and usually lasts from one month to several years. Examples of animals to be administered include livestock such as cows, pigs, horses, and sheep, broilers, hens, quail, ducks, ducks,
Pheasants, poultry such as turkey, yellowtail, red sea bream, blowfish, tuna, flounder, striped horse mackerel, horse mackerel, salmon, carp, eel,
Examples include cultivated aquatic animals such as rainbow trout, sweetfish, shrimp (shrimp, button shrimp, lobster, black tiger, etc.) and crabs (red king crab, snow crab, blue crab, injured crab, etc.).

When the preparation of the present invention is added to feed, the number of bacterial cells is about 10 ⁴ or more per 1 g of feed, preferably about 10 ⁴
It is added so as to be about 10 ⁷ pieces, and more preferably about 10 ⁴ to 10 ⁶ pieces. For example, when a formulation containing no excipient is used, the amount added is 0.1 ppm or more, preferably 0.1 to 1000 ppm, more preferably 0.1 to 1 ppm.
00 ppm is sufficient. When a preparation containing an excipient is used, it may be added in an amount of 0.001% by weight or more, preferably 0.01% by weight to 10% by weight, more preferably 0.1% by weight.
It may be 1% by weight to 0.5% by weight.

When the preparation of the present invention is added to drinking water, the number of cells is about 10 ⁴ or more, preferably about 10 cells per 1 ml of drinking water.
^It is added so as to be ⁴ to 10 ^7, more preferably about 10 ⁴ to 10 ⁶ . For example, when using a formulation containing no excipients, the amount added should be 0.05 ppm or more,
It is preferably 0.05 to 500 ppm, more preferably about 0.05 to 50 ppm. When using a formulation containing excipients, the amount added is 0.0005%.
The above is preferably 0.005% to 5%, more preferably about 0.05% to 0.5%.

Next, the operation of the present invention will be shown by experimental examples. Experimental Example 1-1 Growth test of rainbow trout Purpose: C. butyricum
The dead cells of MIYAAIR were added to the feed, and the following experiment was conducted to examine the growth promoting effect. Experimental method Specimen: C. A feed was prepared by adding 10 ⁵ dead cells of butyricum MIYAIRI / g. Experimental procedure: A rainbow trout having an average body weight of 20 g per animal was bred for 40 days for 525 animals per division. The test results are shown in Table 1.

[0023]

[Table 1] Table 1 Day-to-day growth test of rainbow trout Weight increase (g / tail / day) Feed efficiency Dead cell administration group 0.23 (110) 0.73 (106) Live cell administration group 0.21 (100) 0.69 (100) vs. 0.21 (100) 0.69 (100) ────────────────────────────── ───────────────────────────────────────────────────────────── rate each index As shown in Table 1, C. butyricum MIYA
It was found that dead cells of IRI have a growth promoting effect on rainbow trout.

Experimental Example 1-2 Mouse Growth Test Purpose: C. butyricum MIY
The dead cells of AIRI were added to the feed, and the following experiment was conducted to examine the growth promoting effect. EUT: C. Butyricum MIYAIRI dead cells were added at 10 ⁵ cells / g and 10 ⁷ cells / g to prepare a feed. Experimental procedure: 10 SPF mice each having an average body weight of 25 g were applied to 1 group for 28 days. The test results are shown in Table 2.

[Table 2] Table 2 Mouse growth test Average weight gain group 2.95 g (100) 10 ⁵ dead cells / g administration group 3.80 (129) 10 ⁷ dead cells / g administration group 4 .50 (153) ───────────────────────────────── () is an index with the control section as 100 It was As shown in Table 2, C. butyricum MIYA
It was found that killed cells of IRI have a growth promoting effect on mice.

Experimental Example 1-3 Growth test of broiler Purpose: C. butyri
The killed cells of cum MIYAIRI were added to the feed, and the following experiment was conducted to examine the growth promoting effect. Experimental method Specimen: C. A feed containing 10 ⁶ dead cells of butyricum MIYAIRI / g was prepared. Experimental procedure: A broiler with an average body weight of 40 g was bred for 20 days per group for 28 days. The test results are shown in Table 3.

[Table 3] Table 3 Growth test of broiler control group killed cells average weight gain 882 g (104) 846 g (100) Feed conversion rate 1.57 (98) 1.61 (100) ─────── ─────────────────────────── () is shown as an index with the control plot as 100. As shown in Table 3, C. butyricum MIYA
It was found that killed cells of IRI have a growth promoting effect on broilers.

Experimental Example 1-4 Degradation test of eel feed (dissipation test into water) Purpose: C. butyricum MIYAI
The following tests were carried out in order to check the presence or absence of deterioration by adding live cells and dead cells of RI. Test method 100 g of the feed shown in Table 4 was placed in a beaker, 1.2 times the amount of water was added, and the mixture was kneaded into a sticky form. Then, the mixture was allowed to stand at 30 ° C. for 10 minutes, and 10 g of a lump-like lump was formed to obtain a sample. N. C. butyricum MIYAIRI is 1g sample
It contains 10 ⁵ equivalents.

[0027]

[Table 4] Table 4 Sample composition Control group Viable bacterial cell Dead bacterial cell Fish meal 80% 80% 80% Pregelatinized starch 20 19 19 Live bacteria-1-Death bacteria--1 ──────────── ───────────────────── Place each sample in a 300 ml Erlenmeyer flask and add 100 parts of water.
ml was added, and the mixture was shaken on a heating type shaker for 30 minutes. After shaking, the mixture was left standing for 5 minutes, the supernatant was collected, and the transmittance was measured with a spectrophotometer (wavelength 660 mμ). Each sample was measured 5 times and the average value was calculated.

The test results are shown in Table 5.

[Table 5] Table 5 Deterioration test of eel feed (dissipation test in water) Control group Viable cell Dead cell group Permeability% 56.5 31.8 57.6 (100) (56) (102) ──── ─────────────────────────── () is shown as an index with the control plot as 100. As shown in Table 5, it was found that the use of dead bacteria prevented the dissipation of feed in water due to live bacteria.

Experimental Example 2-1 Developmental test of chicken chick Sample: Product of the present invention: Preparation of Example 2-3 (Number of bacteria: 8.7 × 10 ¹⁰⁾
/ G) Control product: trophozoite dead end of reference example (bacteria number: 5 × 10 ¹¹
Number / g) Experimental procedure: Arbor-acre broiler chicks (body weight 44 g) were used in 20 groups (10 males + 10 females) x 3 replicates per group, and 60 cells were used in the above samples to obtain cell concentration (4.4-4). ．
5) Administer the broiler feed for pre-fabrication (without addition of antibiotics and antibacterial agents) added so as to be 10 ⁵ cells / g, and
A weekly growth test was conducted. In addition, the section to which the feed to which the above sample was not added was administered was used as a blank. The results are shown in the table below.

[0030]

[Table 6] Table 6 Broiler growth test ─────────────────────────────────── Test area Weight gain Dietary requirement Rate ────────────────────────────────── Additive-free group 845g (100) 1.85 (100) Control product Group 864g (102) 1.85 (100) Product of the present invention 884g (105) 1.81 (98) ────────────────────────── ───────── As is clear from the above Experimental Examples 1-1 to 1-4, C.I. bu
The dead bacterial cells of tyricum MIYAIRYAIRI have the effect of promoting the growth of livestock, poultry, and cultured aquatic animals, and are effective in the production of highly safe meat and cultured fish. In addition, in producing feed, there is no limitation like a probiotic agent, and even if it is added to a fish feed, it does not accelerate its deterioration, and it exhibits an excellent effect that a probiotic agent does not. Further, as is clear from Experimental Example 2-1, the C. butyricum
The powder formulation of the present invention comprising vegetatively killed MIYAIRI powder and sugar and / or starch is C.I. butyricum M
The growth promoting effect of IYAAIR is superior to that of vegetative dead bacteria powder alone, and it is effective in improving the productivity of livestock products and marine products.

[0031]

The present invention will be described in more detail with reference to the following examples. Example 1-1 Production of powders Rice bran oil dregs 96 g Soybean oil 3 g Miyairi cell powder (dead cells 10 ¹⁰ / g) 1 g The above components are mixed to give a powder.

Example 1-2 Production of Pellets Rice bran oil residue 70 g Wheat flour 28.9 g Miyairi cell powder (dead cells 10 ¹⁰ / g) 1 g Sodium polyacrylate 0.1 g The above ingredients were mixed and pelletized. Pellet by machine.

Example 1-3 Production of tablets 1 part of Miyairi cell powder (dead cell body 10 ¹⁰ / g) and lactose 80 parts
Part, corn starch 20 parts, low-substituted hydroxypropylcellulose 20 parts and polyvinylpyrrolidone 2 parts were added and mixed well, then granulated, and magnesium stearate 0.
Add 5 parts and mix, then tablet.

Example 1-4 Production of fine granules To 1 part of Myria spp. Powder (dead cells 10 ¹⁰ / g), mannitol 85 parts, carboxymethyl cellulose 15 parts and hydroxypropyl cellulose 2.5 parts were added, and the mixture was thoroughly mixed. Mix and granulate to obtain a fine granule. Example 2-1 Clostridium butyricum MIY
Stock strain of AIRI 588 (FERM BP-278
9) was cultivated at 37 ° using the following medium, and after 16 hours,
The culture solution is centrifuged to obtain vegetative cells. Peptone 1% Yeast extract 1% Glucose 1% 100 g of the obtained bacterial cells, 110 g of corn starch, 50 g of lactose, 8 g of calcium carbonate were suspended in 1 L of water, and the suspension was spray-dried at a hot air inlet temperature of 140 to 160 ° C. The powder of the present invention is obtained by instantaneously making a dry powder at an outlet temperature of 70 ° to 95 °.

Example 2-2 300 g of vegetatively killed Mycobacterium leprae bacillus powder, 100 g of lactose, and 600 g of corn starch were mixed with a v-type mixing group to obtain a powdery preparation of the present invention. Example 2-3 5.15 kg of vegetatively killed vegetative cells of Mycobacterium leprae, 2 kg of lactose,
Water was added to 13 kg of corn starch and 1 kg of calcium carbonate to emulsify, and water was further added to make 100 L, and the mixture was spray-dried with a spray dryer to obtain a powdery preparation of the present invention. Example 2-4 2.5 kg of killed vegetative body of Myria spp., 0.5 gk of lactose
g, 7 kg of corn starch, and 90 kg of brewer's yeast are mixed in a drum mixer to obtain a powdery preparation of the present invention.

Example 2-5 5 g of the spray-dried product of Example 1, 85 g of brewer's yeast and 10 g of defatted rice bran were uniformly mixed to obtain the preparation of the present invention. Reference example Clostridium butyricum MIY
Stock strain of AIRI 588 (FERM BP-278
9) was cultivated at 37 ° using the following medium, and after 16 hours,
The culture was centrifuged in the atmosphere to obtain vegetative cells. Peptone 1% Yeast extract 1% Glucose 1% 100 g of the obtained bacterial cells was suspended in 350 ml of water, and this suspension was preliminarily frozen at -30 ° C and then freeze-dried by a conventional method. Dry powder was obtained to obtain vegetative dead bacterial cell powder.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Kazuo Mizuochi 3-26-17 Akamidai, Konosu City, Saitama Prefecture (72) Inventor Takeshi Asano 871-5 Tateishi, Fujioka City, Gunma Prefecture (72) Inventor Ryuichi Murayama Takasaki, Gunma Prefecture 239 Iwahana-cho, Ichihana Nikka AP F-34 (72) Inventor Toshio Kitagi 3274-3 Wakaho Kawada, Nagano City, Nagano Prefecture (72) Inventor Mamoru Tanaka 2440 Tokura, Tokura Town, Hanashina-gun, Nagano Prefecture

Claims (26)

[Claims] 1. A method for promoting growth of an animal, which comprises orally administering to the animal killed cells of a microorganism belonging to the genus Clostridium. 2. The method according to claim 1, wherein the microorganism belonging to the genus Clostridium is a microorganism belonging to the species Clostridium butyricum. 3. The method according to claim 2, wherein the microorganism belonging to the species Clostridium butyricum is Clostridium butyricum miyairi. 4. The method according to claim 1, wherein the dead cells are vegetative dead cells. 5. The method according to claim 1, wherein the dead cells are added to the feed and administered to the animal. 6. The amount of dead cells in the feed is 10 ⁴ as the number of cells.
6. The method according to claim 5, wherein the number is not less than 1 piece / g. 7. The method of claim 1, wherein the animal is livestock, poultry or aquatic animals. 8. A powdered preparation of dead Clostridium spp. Cells containing, as essential components, vegetative dead cells of a microorganism belonging to the genus Clostridium and sugar and / or starch. 9. The preparation according to claim 8, wherein the microorganism belonging to the genus Clostridium belongs to the species Clostridium butyricum. 10. The preparation according to claim 9, wherein the microorganism belonging to the species Clostridium butyricum is Clostridium butyricum Miyairi. 11. The amount of killed vegetative cells is 10 per 1 g of the preparation.
The formulation according to claim 8, which is ⁶ or more. 12. The amount of dead vegetative cells is 10 per 1 g of the preparation.
The formulation of claim 8, which is ⁶ to 10 ¹¹ . 13. The preparation according to claim 8, wherein the sugar is glucose, fructose, sucrose, oligosaccharide or lactose, and the starch is wheat starch, potato starch or corn starch. 14. A vegetatively killed microbial cell 1 of a microorganism belonging to the genus Clostridium, wherein the sugar and / or starch content in the preparation is 1.
The formulation according to claim 8, wherein the sugar is 0.04 to 700 parts by weight and the starch is 0.2 to 700 parts by weight based on parts by weight. 15. (1) Clostridium butyricum miyairi vegetatively dead microbial cells 0.00005 to 80 w / w
%; (2) Sugar 0.001 to 50 w / w%; (3) Starch 0.015 to 90 w / w%; (4) Poorly water-soluble inorganic substance 0.0005 to 33 w / w%;
And (5) Excipients 1 to 99.9 w / w% are contained, and the number of vegetative cells is 10 per 1 g of the preparation.
The formulation according to claim 8, which is ⁶ or more. 16. The preparation according to claim 15, wherein the saccharide is lactose and the poorly water-soluble inorganic substance is calcium carbonate. 17. A method for producing a powdered preparation of killed Clostridium bacteria, which comprises suspending vegetatively killed bacteria cells of a microorganism belonging to the genus Clostridium and sugar and / or starch in water and then spray-drying. 18. An animal growth promoter comprising, as an active ingredient, dead cells of a microorganism belonging to the genus Clostridium. 19. The growth promoting agent according to claim 18, wherein the dead cells are vegetative dead cells. 20. The growth promoting agent according to claim 18, wherein the microorganism belonging to the genus Clostridium is Clostridium butyricum species. 21. The microorganism belonging to the genus Clostridium is Clostridium butyricum miyairi.
8 growth promoters. 22. A composition of killed bacterium belonging to the genus Clostridium, wherein 0.00005 to 80% of killed cell powder of a microorganism belonging to the genus Clostridium, and the rest being a carrier. 23. The composition according to claim 22, wherein the dead cell powder is a vegetative dead cell. 24. The composition according to claim 21, which contains 0.01 to 50% of dead cell powder of a microorganism belonging to the genus Clostridium. 25. The composition of claim 22, wherein the microorganism belonging to the genus Clostridium is Clostridium butyricum sp. 26. The composition according to claim 22, wherein the microorganism belonging to the genus Clostridium is Clostridium butyricum miyairi.