JPH05246864A - Growth promoter for poultry - Google Patents

Abstract

PURPOSE:To provide the subject promoter containing, as active ingredient, microbial cells belonging to Bifidobacterium thermophilum and/or Lactobacillus salivarius. CONSTITUTION:The objective promoter containing, as active ingredient, microbial cells obtained by culture of Bifidobacterium thermophilum and/or Lactobacillus salivarius of esp. avian origin. This promoter is either administered after addition of an additive into a dry or liquid pharmaceutical, or administered after direct addition to a feed, being applicable to poultry such as chickens, quails, guinea fowls, ducks, turkeys, and pigeons. The microorganisms are anaerobically cultured at 30-42 deg.C, and after culture, cooled and separated. The microbial cells thus recovered is dispersed in a phosphate buffer solution or water containing an amino acid followed by lyophilization. In using this promoter, the surface of the microbial cell powder is pref. coated with an enteric base.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a growth promoter for poultry.

[0002]

2. Description of the Related Art Conventionally, an antibacterial substance is generally administered together with a feed as a growth promoting agent for poultry, but it is not satisfactory due to problems such as resistant bacteria and the development of side effects. In recent years, live-acting agents such as lactobacillus, lactococcus, natto bacterium, butyric acid bacterium, and bifidobacteria have been used instead of the antibacterial substance. However, some of these bacterial species are not intestinal indigenous bacteria, and some of the same bacterial species have a range of animal species that can be established due to the property of animal species specificity. Is.

[0003]

The present inventor has conducted extensive research on a method for promoting the growth of poultry, and as a result, Bifidobacterium thermophilum, particularly Bifidobacterium thermofilm and a Lactobacillus derived from birds. Salivarius (Lactobacillus salivari)
The present invention has been completed by discovering that the administration of bacterial cells obtained by culturing Lactobacillus salivarius derived from birds is effective.

Bifidobacterium thermofilm and Lactobacillus salivarius used in the present invention are described in "Bergey's Manual of Systematic Bacteriology Val.
2 1432 (1986) and 1225 (1986) ”.

In the present invention, all strains of the microorganisms belonging to Bifidobacterium thermofilm and Lactobacillus salivarius can be used.

The above-mentioned two kinds of microorganisms are obligate anaerobic bacteria and facultative anaerobic bacteria, and they grow well under anaerobic and microaerobic conditions. Therefore, culture should be performed under anaerobic conditions. I won't. For example, according to a method of adding an appropriate amount of a mixed solution of L-cysteine hydrochloride and ascorbic acid to a medium immediately after heat sterilization and removing dissolved oxygen in the medium, anaerobic culture can be easily performed even in static culture [[Clinical examination 18: 7-16 (1974)]. It is desirable that the culture temperature is about 30 to 42 ° C, the optimum temperature is about 37 ° C, and the pH is about neutral. After the culture is completed, the culture solution is cooled to about 20 ° C,
Next, the cells are separated by continuous centrifugation at about 5 ± 2 ° C and collected. The collected bacterial cells are dispersed in a phosphate buffer containing amino acids such as sodium glutamate and lysine or water, and then freeze-dried. The form of the bacteria to be administered is not particularly limited, such as dry formulation and liquid formulation,
It may be mixed with feed and given.

However, in order that the administered bacteria reach the intestine without being affected by gastric acid, it is desirable to formulate the bacteria in consideration of acid resistance and mix them with the feed.

For this purpose, it is preferable to coat the surface of the bacterial cell powder obtained by freeze-drying as described above with an enteric base material. The enteric-coated base material used for coating the surface of the bacterial cell powder has the film-forming property used in the technical field of formulation and, when orally administered, the content coated only until it reaches the intestinal tract. It may be any substance to be released, and specific examples thereof include synthetic or semi-synthetic polymer substances such as carboxymethyl cellulose, hydroxymethyl cellulose phthalate and Eudragit, and natural products such as shellac.

When coating the surface of the bacterial cell powder with these enteric substances, the enteric substance is dissolved or dispersed in a solvent, and the bacterial cell powder is added to the obtained solution or dispersion to disperse the solution. The dispersion is sprayed by a spray drying method and simultaneously dried to coat the surface of the bifidobacteria cell powder with an enteric substance, or the enteric substance is dissolved or dispersed in a solvent to obtain a solution or The dispersion liquid is supplied into the high-speed air stream through a nozzle, the bacterial cell powder is separately supplied into the high-speed air stream through a nozzle, and the enteric substance solution or dispersion and the bacterial cell powder are contacted in the high-speed air stream. A method is used in which the surface of the bacterial cell powder is coated with an enteric substance and dried to obtain a bacterial cell powder having a surface coated with a film of the enteric substance.

The solvent in which these enteric substances are dissolved or dispersed may vary depending on the enteric substance selected,
Water, alcohols such as methanol, ethanol, propanol and butanol, ketones such as acetone and methyl ethyl ketone, esters such as ethyl acetate and butyl acetate,
Ethers such as dimethyl ether, diethyl ether and tetrahydrofuran, hydrocarbons such as hexane and pentane and the like can be used alone or in admixture.

In any of the above methods, a solution or dispersion of the enteric substance can be added with a suitable plasticizer such as glycerin monofatty acid ester, castor oil, etc., and the enteric substance of the solution or dispersion. Concentration is 5
-50 (w / w)%, preferably 10-32 (w / w)
It is assumed that the concentration is about%.

The apparatus used for producing the bacterial cell powder whose surface is coated with the enteric-coated substrate by the above-mentioned spray drying method may be an ordinary spray dryer. In addition, the device used for the production of the target product by the method of contacting the bacterial cells with the enteric-coated substrate solution or dispersion in a high-speed air stream and the subsequent drying is, for example, Dispacoat manufactured by Nisshin Engineering Co., Ltd.
There is ^R.

According to another embodiment of the present invention, the cells of the above-mentioned microorganism can be granulated or tableted, and the surface of the granule or tablet can be coated with the above-mentioned enteric base material. When granulating or tableting the bacterial cells, a known granulating method or a tableting method using a tableting machine is used. Enteric coating of these granules or tablets may be performed using a coating pan.

Examples of poultry to which the growth promoter of the present invention is applied include chickens, quail, guinea pigs, ducks, turkeys, pigeons and the like.

The growth promoting agent of the present invention is preferably used by oral administration. In this case, this growth promoter may be formulated by adding additives so that it can be easily ingested by poultry itself in the course of formulation, or may be mixed with other feed for administration. .. The present invention will be described below with reference to specific examples.

Example 1 i) Preparation of fungal agent of chicken-derived Bifidobacterium thermofilm chN118 strain Bifidobacterium thermofilm chN118 strain (this strain is a microorganism of the Agency of Industrial Science and Technology on February 18, 1992) An anaerobic (L-cysteine hydrochloride and ascorbine) was used for 18 hours at 37 ° C. using 5 liters of MRS medium (manufactured by OXOID), which was deposited at the Institute of Industrial Science and Technology with a deposit number of Microorganism Research Institute No. 12762). The mixture of the acid is added to 1%), and the mixture is centrifuged (15000 rpm) to collect the cells, and the cells are dispersed in a phosphate buffer containing sodium glutamate and soluble starch, and then lyophilized. After drying, the dry powder was dispersed in a shellac solution (25% ethanol solution), and this dispersion was subjected to surface modification using a powder surface modification device (manufactured by Nisshin Engineering Co., Ltd.) DC-8 type, The acid-resistant bacterial agent adjusted to the concentration was obtained.

Ii) Preparation of a fungal agent of chicken derived Lactobacillus salivarius chN426 strain Lactobacillus salivarius chN426 strain 3) using 5 liters of MRS medium (manufactured by OXOID), which was deposited under the accession number of No. 12763).
Cultivated anaerobically at 7 ° C for 18 hours (1% mixture of L-cysteine hydrochloride and ascorbic acid was added), centrifuge (15000 rpm) to collect cells, and collect sodium glutamate and soluble starch. Disperse in a phosphate buffer containing the product and freeze-dry. After drying, dry powder was dispersed in shellac solution (25% ethanol solution), and this dispersion was used for powder surface modification equipment (Nisshin Engineering
Surface modification was performed using DC-8 type (manufactured by Co., Ltd.) to obtain acid-resistant fungal agents adjusted to various concentrations.

Iii) Preparation of a fungal agent of Lactobacillus acidophilus chN253 strain derived from chicken MR strain of Lactobacillus acidophilus chN253 strain
5 liters of S medium (OXOID) at 37 ° C
It was anaerobically cultured for 8 hours (1% of a mixture of L-cysteine hydrochloride and ascorbic acid was added), and this was centrifuged (15
000r.pm) to collect the bacterial cells, sodium glutamate,
After being dispersed in a phosphate buffer containing soluble starch, it is freeze-dried. After drying, the dry powder was dispersed in shellac solution (25% ethanol solution), and this dispersion was used as a powder surface reforming device (manufactured by Nisshin Engineering Co., Ltd.) DC.
Surface modification was carried out using -8 type to obtain acid-resistant bacterial agents adjusted to various concentrations.

Iv) Preparation of fungal agent of Bifidobacterium thermofilm PgN125 strain derived from swine Bifidobacterium thermofilm PgN125 strain was used at 37 ° C. in 5 liters of MRS medium (manufactured by OXOID).
Cultivated anaerobically for 18 hours (adding 1% of a mixture of L-cysteine hydrochloride and ascorbic acid), centrifuging this (15000 rpm) to collect bacterial cells, and phosphorus containing sodium glutamate and soluble starch. After dispersing in acid buffer, freeze-dry. After drying, dry powder was dispersed in shellac solution (25% ethanol solution), and this dispersion was used for powder surface modification equipment (Nisshin Engineering Co., Ltd.).
(Manufactured by K.K.) DC-8 type was used for surface modification to obtain acid-resistant bacterial agents adjusted to various concentrations.

V) Preparation of fungal agent of pig-derived Lactobacillus acidophilus PgN311 strain Lactobacillus acidophilus PgN311 strain MR
5 liters of S medium (OXOID) at 37 ° C
It was anaerobically cultured for 8 hours (1% of a mixture of L-cysteine hydrochloride and ascorbic acid was added), and this was centrifuged (15
000r.pm) to collect the bacterial cells, sodium glutamate,
After being dispersed in a phosphate buffer containing soluble starch, it is freeze-dried. After drying, the dry powder was dispersed in shellac solution (25% ethanol solution), and this dispersion was used as a powder surface reforming device (manufactured by Nisshin Engineering Co., Ltd.) DC.
Surface modification was carried out using -8 type to obtain acid-resistant bacterial agents adjusted to various concentrations.

Vi) Preparation of fungal agent of Lactobacillus salivarius PgN226 strain derived from pig Lactobacillus salivarius PgN226 strain MRS
Use 5 liters of culture medium (manufactured by OXOID) at 37 ° C for 18
The culture was performed anaerobically (addition of 1% of a mixture of L-cysteine hydrochloride and ascorbic acid), followed by centrifugation (150).
The cells are collected at 00 rpm.pm, dispersed in a phosphate buffer containing sodium glutamate and soluble starch, and then lyophilized. After drying, the dry powder was dispersed in shellac solution (25% ethanol solution), and this dispersion was used as a powder surface reforming device (manufactured by Nisshin Engineering Co., Ltd.) DC.
Surface modification was carried out using -8 type to obtain acid-resistant bacterial agents adjusted to various concentrations.

Vii) Administration of fungal agent Broilers (harvard) of 0-day-old were divided into 7 groups with 20 birds per group, and 1 group was used as a control (group without added fungal agent).
The remaining 6 groups were treated with fungal agents, and 10 ⁸ fungal agents of each of the bacterial species shown in Table 1 prepared above were orally administered once a day for 1 week from the age of 0 days after birth. Administered, 3
The weight gain and the feed requirement rate [the amount of feed (kg) required to increase the body weight by 1 kg] after the week were determined. As shown in Table 1, chicken-derived Bifidobacterium thermofilm chN1
18 strains and Lactobacillus salivarius chN42
The effect was recognized in 6 strains.

[0023]

[Table 1]

Example 2 A broiler (harvard) aged 0 days was divided into 8 groups with 20 birds per group for one test, and 1 group was used as a control (group without added fungal agent), and the remaining 7 groups. Is a group to which the fungal agent is administered (all of the used bacterial strains are chicken-derived strains), and the bacterial agent of each bacterial strain shown in Table 2 prepared in Example 1 is used for 10 ⁵ cells / day per bird.
0 ⁶ and 10 ⁸ were orally administered once daily for 1 week from the age of 0 days after birth, and the weight gain and the feed conversion rate were determined after 3 weeks. As shown in Table 2, the effect was observed on the Bifidobacterium thermofilm chN118 strain, the Lactobacillus salivarius chN426 strain, and a mixed group of these two bacterial species. Particularly in the mixed group, the effect was specifically observed even in the 10 ⁶ group.

[0025]

[Table 2]

Example 3 Field application test A group of 320 days old broilers (Harvard)
Feathers (80 feathers x 4 pens) were divided into 5 groups, and 1 group was used as a control (no-addition group), and the remaining 4 groups were used as fungal agent administration groups. The amount of each fungal agent added is Bifidobacterium per ton of feed.
The thermofilm chN118 strain group is 5 × 10 ¹⁰ and the Lactobacillus salivarius chN426 strain group is 5 × 10 ^9.
The concentration of Bifidobacterium thermofilm ch
N118 strain and Lactobacillus salivarius chN
The total number of bacteria in the mixture of 426 strains was 5.5 × 10 ¹⁰ and 1.1 × 1.
Two groups with a concentration of 0 ¹¹ were set up.

The results obtained are shown in Table 3. From this table, it was confirmed that the body weights of the two groups of the mixed administration group were continuously increased significantly after the age of 30 days and the feed conversion rate was improved as compared with the single administration group.

[0028]

[Table 3]

[0029]

[Table 4]

Claims (1)

[Claims] 1. A growth promoting agent for poultry, which comprises Bifidobacterium thermofilm or Lactobacillus salivarius cells or a mixture of both as an active ingredient.