JPS59179037A - Feed additive and feed for animal - Google Patents

Abstract

PURPOSE:To provide a feed additive and feed for animal, containing the cells of Bacillus licheniformis, free from biotoxicity, and effective in increasing the body weight of aminals. CONSTITUTION:A feed additive or feed for animal containing the cells of Bacillus licheniformis as active component. The Bacillus licheniformis is a facultative anaerobe belonging to Bacillus genus, and the mutant of the strain can be used effectively as well as the strain separated from the natural environment. Preferable strain is a mutant, Bacillus licheniformis MN-001 (FERM-BP No.266). The Bacillus licheniformis strain can be cultured by conventional fermentation process in liquid medium or solid medium. Any medium such as natural medium, semisynthetic medium, synthetic medium, etc. can be used for the purpose.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel animal feed additives and feeds. More specifically, the present invention relates to animal feed additives and feeds containing /4 Bacillus licheniformis as an effective bacterium.

Currently, animal feed is changing from mainly homemade feed to compound feed produced in modern factories.
The ratio of compound feed to total feed is extremely high.

In addition to the main ingredients of grains, oil cake, fishmeal, and fats and oils, these compounded feeds also contain amino acids, minerals, vitamins, and antibiotics for various purposes such as nutritional supplementation, improving digestion and absorption, promoting growth, and preventing diseases. Substances, enzymes, probiotics, etc. are used as auxiliary raw materials.

However, among these antibiotics, restrictions on their use have been tightened due to the risk of residual migration causing allergies and changes in the fetal flora in humans.
The range of use is being limited or reduced.

As enzymes, glucose agents, amylase agents, cellulase agents, protease agents, etc., and mixtures of these enzyme agents, etc., are being considered, but the current situation is that their effects vary and a constant effect cannot always be expected. It is.

In addition, Bacillus natto, lactic acid bacteria, butyric acid bacteria, bifidobacteria, and the like have been studied as viable bacteria agents, and although some effects have been seen, they are still not sufficient.

As seen above, the reality is that there is still no satisfactory animal feed additive.

In view of this situation, the present inventors investigated various types of Bacillus bacteria isolated from nature that are effective as animal feed additives, and found that Bacillus licheniformis is not toxic to animals and is not toxic to animals. However, it was confirmed that there was a considerable increase in weight, and the present invention was completed.

The present invention is an animal feed additive and feed characterized by containing Bacillus licheniformis as an effective bacterium.

Bacillus licheniformis used in the present invention is a facultative anaerobic bacterium belonging to the genus Bacillus, and not only bacteria isolated from nature but also mutant strains thereof can be effectively used. Among them, a preferred strain is MN-
This is the mutant strain Bacillus licheniformis MN-001 deposited as 001.

This Bacillus licheniformis strain MN-001 (hereinafter M
It is designated as the N-001 strain. ) shows the mycological properties of

A, form (1) Bacillus with a width of 0.5 to 08 μm and a length of 2 to 6 μm.

(2) Durham staining: Positive.

(3) Forms oval endospores.

(4) Motility: Yes.

B. Growth (1) Ordinary agar medium: Very good growth. No pigment formation.

(2) Medium mainly composed of molasses and ammonium sulfate: Grows very well. No pigment formation.

(3) Salt liquid medium Section 5: Grow.

7chi: Grow.

10%: Grows.

C0 Physiological properties (1) Catalase production 10 (2) Growth under anaerobic conditions 10 (31VP reaction + (4) Growth condition at high temperature 50℃ + 55℃ - (5) Presence or absence of acid production from sugars Glucose + L-arabinose + D-xylose + mannitol + D-tagatose + (6) Arginine dihydrolase + (7)
Lecithiner 7 - (8) Hydrolyzability of starch + (9) Hydrolyzability of gelatin
1α0) DNA hydrolyzability + side Citrate availability 1゛(1,2+ Propionate availability 1α□□
□ Nitrate reducing property 1αa Indole production - (Deamination of 151 phenylalanine 1) To culture Bacillus licheniformis, which is the effective bacterium of the present invention, liquid culture or solid culture is performed according to the usual fermentation method. The medium can be any natural medium, semi-synthetic medium, synthetic medium, etc.The composition of the medium is as follows: (1) As a carbon source, glucose, lactose, dextrin/glycerin, Starch, molasses, etc. (II) Nitrogen sources include starch, meat extract, yeast extract, dried yeast, soybean meal, urea, thiourea, ammonium salts, nitrates, and other organic or inorganic nitrogen compounds as 010 inorganic salts. 7. Magnesium, manganese, potassium, calcium,
Phosphates, nitrates, carbonates, chlorides, etc. of iron etc. are used. Amino acids, vitamins, nucleic acids and related compounds may also be added. The culture temperature is based on the usual fermentation method, but the culture can be suitably carried out at, for example, 20°C to 50°C. The culture period is also similar to the usual fermentation method, but for example, 12 hours to 5 days is suitable for expansion.

The culture thus obtained can be used as it is, or by washing repeatedly and using only the bacterial cells. It can be used as a feed additive or as a feed after being added to the preparation, freeze-dried, spray-dried, etc.

In general, when used as animal feed or feed additives, for example, the culture solution may be used as is, or wheat flour, starch, dextrin, etc., grains used as raw materials for feed, grains such as defatted rice bran, oil cake, etc. It is sufficient to form a formulation using an appropriate diluent.Also, it is also possible to wash and separate only the bacterial cells from the culture solution, formulate the same formulation, and use it.

The Bacillus licheniformis live bacterial preparation obtained in this way can be directly mixed into animal feed or used as a feed additive for beef cattle, dairy cattle, calves, pigs, piglets, sheep, wild animals, and horses. ,
For domestic animals such as rabbits, dogs, and cats, and for poultry such as meat chickens, egg-laying hens, chicks, breeding hens, geese, turkeys, quail, and foals, etc.] Providing lentils to increase weight and improve health. It is valid.

The dosage also depends on the target animal and the age of the target animal.
In addition, -elK cannot be specified as it may vary depending on other ingredients of the feed, but normally the number of live Bacillus licheniformis bodies per edge of the feed is 102 to 1015, preferably 1.
It is preferable to add 0' to 10'2 pieces, most preferably 105 to 1010 pieces.

Next, production examples and examples of the present invention will be shown.

Production example [Preparation of Bacillus licheniformis live bacteria preparation] Tap water 100%! 2 parts by weight of sugar, 1 part by weight of ammonium sulfate, 0.1 parts by weight of dibasic phosphoric acid, 0 parts by weight of yeast extract.
．． Dissolve two parts of 0.05 heavy duty. One ton of culture solution whose pH was adjusted to 80 with IN hydrochloric acid and 0.1N force neutralizer was placed in a jar fermenter, and after killing (121°C, 15 minutes), precultured Bacillus licheniformis strain MN-001 (minimal MN-001 on Koken Joyori No. 266
and U7 (deposited) were inoculated and cultured for 24 hours. Next, the bacterial cells were separated from the culture solution by centrifugation, and starch I Ky was added to and mixed with the bacterial cells, and then dried to obtain Bacillus licheniformis MN-[101 strain 1AIJ (
(to) MN-001 formulation. ) was obtained. The number of viable bacteria in this preparation was 106/1.

In addition, a formulation was also made using defatted rice bran instead of starch.

Example 1 (Breeding of broilers) Using 120 broilers (60 each of males and females), 40 in each group.
The birds were divided into 6 groups (20 males and 20 females), and the MN obtained in the production example was fed to the basic feed shown in Table 1, a control group (starch added group), and a control group (starch added group).
A group containing -001jM agent (containing 108 bacteria per 11 starches) was established, and a feeding test was conducted for 8 weeks. The results are shown in Table 2.

Table 1 (Composition of basic feed) (Note) Standard test feed manufactured by Nippon Compound Feed @SD broiler early stage* 8D broiler first stage** Crude protein ≧ 2
6 ≧ 19 Crude fat ≧ 5
≧ 6 coarse fiber ≦ 4 ≦ 4 coarse fiber ≦ 6.5 ≦ 685ME
(Kaxshi49) ≧3000
≧6100 Table 2 Example 2 (Piglet Rearing) Thirty weaned piglets (15 of each sex) approximately 25 days old were used and divided into 6 groups of #10 piglets (5 of each sex). In each group, the basic feed shown in Table 6 was added to the control group (defatted rice bran added group) and the natto bacteria preparation "Glogen 8" (trademark, manufactured by Eisai@, containing io8 natto bacteria per starch). Ku, MN-001 formulation (preparation containing 1?10' of live bacteria obtained using defatted rice bran in the manufacturing example) added needle 6
A 14-week feeding test was conducted in a separate section. 4th result
It is shown in the table.

Table 6 (Basic feed composition) Figures are based on the weight part.
The preparation request rate and digestibility can be significantly improved.

Procedural Amendment February 1982 114. Mr. Commissioner of the Japan Patent Office 1. Indication of the case % Application No. 58-051482 2. Name of the invention Animal feed additives and feed 3. Person making the amendment Relationship to the case Patent applicant address Kasumigama Pressure, Chiyoda-ku, Tokyo Chome 2-5 Name (312) Mitsui Toatsu Chemical Co., Ltd. Representative Kasama
Yubebe, 4th generation Osamu Address: 19-14-6, Toranomon-chome, Minato-ku, Tokyo 105 Contents of the specification subject to amendment Zm (1) "Significant weight gain" on page 6, line 12 of the specification is corrected to "various effectiveness."

(2) Insert the following sentence next to the bottom line of page 12 of the specification.

"Example 6 Using 4Ω pregnant sows weighing 170-180kl, they were divided into two groups with LO pigs in each group. ), control group (starch added group), MN-00
1 preparation (108 per 1 g obtained using starch in the production example)
A test group was set up with the addition of a preparation containing several viable bacteria, and a feeding test was conducted for more than 80 days, from 60 days before calving to 21 days after calving. The results are shown in Table 2.

Table 1 Components of Hibreed B Crude protein 150% or more Crude fat
2.0% or more crude fiber io,
o % or less Crude ash 10.0% or less Calcium 0.6% or more Phosphorus
0.45% or more digestible crude protein 11.5% or more total digestible nutrients
72.0% or more Table 2 Using 10 1-week-old Holstein male calves, divide them into 2 groups of 5 cows each, and add starch to the basic milk replacer with the composition shown in Table 1 for each group. Control milk replacer and MN-
001 formulation (91 per gram obtained using starch in the production example)
A milk replacer for the test plot was prepared by adding a preparation containing 0.08 viable bacteria) and used as a control plot and a test plot, respectively. In the feeding test, 1 part by weight of the milk replacer was dissolved and dispersed in 7 parts by weight of warm water and then fed to calves for a 4-week test.

All results are shown in Table 2.

In addition, in recent methods of feeding calves in Japan, a method of feeding whole calf feed (lower in protein and fat than milk substitute) called calf starter along with milk replacer as mentioned above has been widely adopted. In this example, the calf starter was not used in combination because the purpose was to fully examine the effects of the milk replacer itself.

Table 1 Note 1) Contains vitamins and minerals Table 2 Note 2) Milk replacer (powder equivalent value) Example 5 68 weeks ◆ From 100 White Leghorn chickens, 60 hens with high egg-laying rates over the previous 4 weeks were used. It was selected and divided into two categories. A control group (starch-added group), MN-001 formulation (produced using starch in the manufacturing example, After a 1-week preliminary breeding period, a 2-week feeding test was conducted.The results are shown in Table 2.

Table 1 Ingredients of Mylayer 17 Crude protein 17% or more Crude fat 6.5% or more Calcium 2.8% or more Phosphorus
055% or more crude fiber
5.0% or less Coarse ash content 1 ro or less

Claims (1)

[Claims] 1. Bacillus licheniformis
1. An animal feed additive characterized by containing P. licheniformis as an effective bacterium. 2. Bacillus licheniformis bacterium
Claim 1, which is the Bacillus licheniformis strain MN-001 deposited as MN-001 in No.
Animal feed additives listed in Section 1. 6. An animal feed characterized by containing Bacillus licheniformis as an effective bacterium. 4. Bacillus licheniformis bacterium
The animal feed according to claim 6, which is the Bacillus licheniformis strain MN-001 deposited as MN-001 in