MXPA99011167A - Method for administering viable microorganism composition for poultry - Google Patents

Abstract

A method for administering viable microorganism compositions for poultry, comprising:administering a viable microorganism composition comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii at the stage of newborn fledgling;and administering a viable microorganism composition which comprises a viable microorganism belonging to Bacillus subtilis.

Description

i / IODOD FOR ADM I N ISTRAR A N O COM POSITION OF I ICROORGAN VIABLE ISMS FOR BIRDS OF CORRAL TECHNICAL FIELD The present invention relates to a method for administering viable microorganism compositions to poultry, in which compositions of viable microorganisms useful for the growth of poultry are administered, thereby feeding the poultry with a high productivity, while reducing the quantities of drugs, such as antibiotics, antibacterial agents and the like, or without the administration of the drugs.

BACKGROUND ART In the feeding of poultry, such as young chickens and the like, the dense breeding of 40 to 60 birds per 1 tsubo (ca. 3.3 m2) of a poultry farm is generally carried out for the purpose to reduce the cost of production. Additionally, poultry are fed to achieve rapid growth, 40 to 55 g in average daily body weight gain, so that birds can be shipped about 2 months after incubation. Under these conditions, poultry are always under stress, and often get sick, and diseases spread among poultry. In floor feeding, because poultry stings food and the like, which are smeared with excrement on the floor, toxic bacteria spread very easily over all birds in the feeding facility. In addition, in an open power installation, or even in a windowless power installation in some cases, toxic bacteria can be easily spread through the power installation. In addition, for example, although a young chicken house is disinfected each time the young chickens are shipped, because smaller animals, such as rats and the like, in the surrounding area are not affected by this treatment, since these animals they carry bacteria, such as salmonella and the like, to the poultry house, when fresh chickens are introduced into the feeding facility again there is a latent contamination. In order to maintain a high productivity while avoiding a decrease in the speed of growth and similar during the feeding period, and also to avoid contamination of poultry with toxic bacteria under these conditions, several drugs, such as, antibiotics, antibacterial agents and the like, are generally administered to poultry. However, due to this, there is the possibility of residual drugs in meat and eggs, and also the danger of generating bacterial strains resistant to drugs (as has now been reported in many cases), which has increased the demand between users for drug-free livestock farm products, produced without using medications. On the other hand, under natural conditions without dense breeding of a large number of birds, high productivity can not be expected. However, it is possible to raise poultry, such as chickens and the like, without using medicaments, such as antibiotics, antibacterial agents and the like. The meat and eggs of chickens raised under natural conditions, namely the so-called natural farmyard chickens, are now in great demand due to safety and good taste, and are traded in the market at a high price. Accordingly, great interest has been directed towards the development of a feeding method for raising poultry, such as chickens and the like, without causing contamination with toxic bacteria, while maintaining high productivity, by a method of drug-free feeding (feeding in which drugs, such as antibiotics, antibacterial agents and the like, are not used during part of, or throughout, the feeding period, so that the drugs do not remain in the birds, at least at the time of shipment). When the conventional dense breeding of a large number of poultry is done by simply avoiding the administration of medicines, the poultry are easily infected with toxic bacteria. Consequently, it is not possible to maintain a productivity at a commercially acceptable level without causing contamination with toxic bacteria. As a means to reduce the infection of poultry with toxic bacteria without relying on medicaments, such as antibiotics, antibacterial agents and the like, it has been proposed to administer microorganisms and the like, which are useful for the growth of several poultry. Bacteria belonging to the genus Bacillus, lactic acid bacteria belonging to the lactobacillus genus, bacteria belonging to the genus Bifidobacterium and the like are known as useful microorganisms for food additives or as various preparations of EC (competitive exclusion). Some of these microorganisms are commercially available as preparations of viable microorganisms from food additives for poultry. However, although the effects are recognized to some degree, they are not sufficient. In particular, no means has been reported, which can perform the free feeding of medicines of poultry in dense breeding, with a large number of poultry, and with productivity at a commercially acceptable level by providing these preparations of viable microorganisms. For example, Japanese Registered Patent No. 2528055 and JP-B-3-79988 (the term "JP-B" as used herein means a "examined Japanese Patent Publication") describe that the profit ratios can be improved. of body weight and conversion of feed into animals, and that effects, such as controlling action of intestinal function and the like, can be obtained by administering a preparation of viable microorganisms comprising Bacillus subtilis to animals. It is possible to perform a drug-free diet by applying the medium to, for example, young chickens, but only in certain limited regions where young chickens are not produced on a large scale. However, in many regions where chickens are often fed, because they are frequently infected with toxic bacteria, which are easily spread through the feeding facilities as described above, commercial feeding is extremely difficult. drug free of young chickens using this medium. In addition, even if it were possible to feed free medicines of young chickens using this medium, the proportion of body weight gain of poultry is slower than feeding using drugs. As a result, the high productivity of the diet with applied drugs is not obtained. In addition, preparations of viable microorganisms currently on the market have problems, such as, the reduction of the viable account during the distribution steps, the poor colonization capacity after the formation of the intestinal bacterial flora, the need for continued administration of the preparations of viable microorganisms for a prolonged period, and the like. Particularly, the need for the continued administration of the viable microorganism preparations over a prolonged period, due to the poor colonization capacity after the formation of the intestinal bacterial flora, is currently a serious obstacle to practical use from the economic point of view .

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a method for administering viable microorganism compositions to poultry, so that free feeding of poultry medications can be performed while maintaining good productivity.

To solve the aforementioned problems, the inventors of the present invention have performed the classification of useful bacteria from broad natural sources, and have found that certain lactic acid bacteria isolated from the intestinal tract of a chicken can be easily colonized in the tract of newborn chickens in a short time after incubation. In addition, the inventors have found that, in particular, a heterofermentation lactic acid bacteria isolated from the ileum of a chicken and homofermentation lactic acid bacteria isolated from the caecum of a chicken can be easily colonized in the intestinal tract of chicks and have several effects, such as, inhibition of growth of toxic bacteria, prevention of diarrhea, intensification of growth, improvement of growth rate, improvement of feed conversion ratios and the like. In addition, the inventors have found that lactic acid bacteria can be colonized in the intestinal tract of newborn chickens as soon as they are born, by simply atomizing a suspension of viable microorganisms from these Lactobacillus lactic acid bacteria once to freshly hatched chickens. born within 4 days after hatching. Additionally, the inventors have found that, when useful lactic acid bacteria are colonized in combination with the administration of viable Bacillus subtilis microorganisms during a period from the chick stage to the mature bird stage in the poultry free feed. of barnyard, the body weight gain can be improved in the initial feeding stage and a sufficient growth rate can be obtained in comparison with the simple administration of viable Bacillus subtilis microorganisms, so that it becomes possible to perform the drug-free feeding of poultry through dense breeding in a large number of poultry with high commercially effective productivity. The present invention has been achieved on the basis of these findings. Thus, the present invention relates to a method for administering viable microorganisms to poultry, comprising: administering a first composition to a poultry comprising viable bacteria that belong to Lactobacillus reuteri and Lactobacillus johnsonii, in the stage of newborn chicks. Furthermore, the present invention relates to a method for administering viable microorganism compositions for poultry, comprising: administering a composition of viable microorganisms for poultry comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii in the stage of newborn chicks; and administering a viable microorganism composition for poultry, comprising a viable microorganism belonging to Bacillus subtilis.

BEST MODE FOR CARRYING OUT THE INVENTION The method of the present invention for administering viable microorganism compositions for poultry includes a composition of viable microorganisms for poultry comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii (from hereinafter frequently referred to as a "composition of viable microorganisms of lactic acid bacteria"), which is administered to the newborn chicks. The lactic acid bacteria described above belonging to the genus Lactobacillus are facultative anaerobic bacteria. They can be used effectively not only those isolated from the intestinal tract of poultry, but also other strains isolated from a natural source. Preferred examples of the strain belonging to Lactobacillus reuteri include Lactobacillus reuteri CP-720 (Deposit No. FERM BP-6332), Lactobacillus reuteri CP-722 (Deposit No. FERM BP-6334) and the like. The preferred example of the strain belonging to Lactobacillus johnsonii includes Lactobacillus johnsonii CP-721 (Deposit No. FERM BP-6333) and the like. These three strains have been deposited in the National Institute of Bioscience and Human Technology, Agency for Industrial Science and Technology (Address: 1 -3, Higashi 1 Chome, Tsukuba-shi, Ibaraki-ken 305-8566, Japan) April 27, 1998. Other strains of Lactobacillus reuteri and Lactobacillus johnsonii are known, such as those described in A TCC Bacteria and Bacteriophages, 19th ed. , 1996, pages 1 95 and 1 97, incorporated herein by reference. The bacteriological properties of Lactobacillus reuteri CP-720 and CP-722 and Lactobacillus johnsonii CP-721 are shown in Table 1.

Table 1 The composition of viable microorganisms of lactic acid bacteria can also contain heterofermentation lactic acid bacteria belonging to the genus Lactobacillus other than Lactobacillus reuteri, such as Lactobacillus brevis, Lactobacillus buchneri and the like, and also lactic acid bacteria of homofermentation belonging to the genus. Lactobacillus other than Lactobacillus johnsonii, such as, Lactobacillus gasseri, Lactobacillus crispatus and the like, belonging to the Lactobacillus acidophillus group. Many different strains of many different genera of Lactobacillus are known, such as those described in A TCC Bacteria and Bacteriophages, 19th ed. , 1 996, pages 1 92-1 99, incorporated herein by reference. Preferred examples of the medium, which can be used to culture viable microorganisms of lactic acid bacteria, include milk medium, such as cow's milk, goat's milk, horse's milk and similar, skimmed milk thereof and a medium for lactic acid bacteria, such as BL medium, Briggs liver broth medium, MRS medium, GAM medium, TTY medium and similar. The lactic acid bacteria can be grown at 25 to 45 ° C, more preferably 30 to 40 ° C, and for 6 to 30 hours, more preferably 10 to 24 hours. The culture broth thus obtained can be used directly as the composition of viable microorganisms of lactic acid bacteria by storage as such at about 5 ° C until use. Alternatively, the microorganisms can be recovered by centrifugation, mixed with a protective agent and then lyophilized under vacuum. The resulting powder of the bacteria can be stored in a cold, dark chamber and used as a viable microorganism composition of lactic acid bacteria, by suspending, mixing or dissolving the powder when used. The dry microorganism powder prepared in this form is more preferred because it can withstand long periods of storage. The composition of viable microorganisms of lactic acid bacteria can contain a carrier and diluent. The carrier and diluent are not particularly limited, and are selected from pharmaceutically or nutritionally acceptable carriers and diluents. In addition, the composition of viable microorganisms of lactic acid bacteria can be contained in a poultry feed (ration). In the present invention, microorganisms belonging to Lactobacillus reuteri and Lactobacillus johnsonii can be subjected to an appropriate mutation treatment, such as exposure to ultraviolet light, X-rays or radiation, and a chemical treatment with a mutagenic compound (e.g. acridine dye, nitrosoguanidine). Mutants can also be prepared by insertion, deletion or substitution of nucleotides, as well as spontaneous mutation. The terms lactobacillus reuteri and Lactobacillus johnsonii include these mutants. The term "newborn pigeon stage" means just after hatching of the pigeon (eg, chicken, pigeon, chick or the like), specifically, a period of from 0 to about 4 days after the exit of the pigeon. shell The time for administration of the viable microorganism composition of lactic acid bacteria is not particularly limited, as long as and during the newborn pigeon stage; however, it is preferred to administer the composition within 4 days after hatching, most preferably within 2 days after hatching, a period prior to colonization of intestinal bacterial flora in the intestinal tract of the pigeon . The number of times of administration is not particularly limited; however, sufficient effects can be obtained by a simple administration. The effects obtained become more stable and secure as the number of times of administration increases. Nevertheless, the administration of three times or more is not economic, and the effects obtained with it are almost the same as with a two-time administration. Although not particularly limited, administration of the viable microorganism composition of lactic acid bacteria can be accomplished by oral administration. Specifically, the composition can be administered orally by adding it to drinking water or the like, and allowing the chicks to freely swallow it, or by spraying it from a position above the chicks using an atomizer or the like. The method of administration with a lifting atom is preferred due to the habit of chirping chicks while opening their peaks upwards, so that the composition of viable microorganisms can be administered orally having a high concentration in the stage of newborn chicks easily and easily. safe. To perform the administration, the total density of lactic acid bacteria in the composition of viable microorganisms of lactic acid bacteria is, preferably, from 10 6 to 10 010 viable microorganisms per g, more preferably, from 10 7 to 10 9 viable microorganisms. by g. In addition, the composition of viable microorganisms of lactic acid bacteria is preferably administered in an amount of 1 x 10 3 to 1 x 10 1 or viable microorganisms per pigeon, more preferably 1 x 1 04 to 1 x 1 06 viable microorganisms per pigeon. In the method of the present invention, a composition of viable microorganisms for poultry comprising a viable microorganism belonging to Bacillus subtilis (hereinafter often referred to as a "viable microorganism composition of Bacillus subtilis") is administered in addition to the administration described above of the composition of viable microorganisms of lactic acid bacteria. Preferred examples of the Bacillus subtilis strain include Bacillus subtilis C-3102 (Deposit No. FERM BP-1 096), which has been deposited with the Institute of Human Bioscience and Technology (former name: Fermentation Research Institute) , Industrial Science and Technology Agency (address: 1 -3, Higashi 1 chome, Tsukuba-shi (old address: Yatabe-machi, Tsukuga-gun), Ibaraki-ken 305-8566 (old zip code: 305), Japan) on June 28, 1986, and the like. The bacteriological properties of Bacillus subtilis C-31 02 are already described in Japanese patent no. 2528055, JP-B-3-79988 and US Pat. No. 34,837. Many different strains of Bacillus subtilis are known, such as those described in A TCC Bacteria and Bacteriophages, 19th ed. , 1996, pages 57-63, incorporated herein by reference. To cultivate Bacillus subtilis C-3102, an aqueous or solid medium containing materials, such as carbon sources, nitrogen sources, inorganic substances and the like, which are generally used to culture microorganisms, can be used as the culture medium. Examples of carbon sources include those that can be assimilated, such as glucose, fructose, sucrose, starch, molasses and the like. Examples of nitrogen sources include peptone, meat extract, casein hydrolyzate, ammonium sulfate and the like. As the occasion demands, phosphates, magnesium salts, postasium, sodium, calcium, iron, manganese and the like, vitamins, amino acids, antifoaming agents, surfactants and the like, may also be added as inorganic components. The cultivation is preferably done aerobically. The initial pH of the medium is preferably 5 to 9, more preferably 6 to 8; the culture temperature is preferably 20 to 50 ° C, more preferably 35 to 40 ° C; and the culture period is preferably 12 hours to 7 days. The culture mixture obtained in this manner can be used as the viable microorganism composition of Bacillus subtilis, as such, or as its concentrated product or as cells isolated therefrom, directly or after adding additives, such as fillers and the like. . The fillers are not particularly limited, and examples include calcium carbonate, defatted rice bran, corn kernels, corn flour, wheat bran, skimmed milk powder and the like. The viable microorganism composition of Bacillus subtilis can be contained in a carrier or diluent. The carrier and diluent are not particularly limited, and are selected from pharmaceutically or nutritionally acceptable carriers and diluents. In addition, the viable microorganism composition of Bacillus subtilis can be contained in a poultry feed. In the present invention, the microorganism belonging to Bacillus subtilis can be subjected to a suitable mutation treatment, such as exposure to ultraviolet light, X-rays or radiation, and a chemical treatment with a mitogenic compound (for example, acridine dye). , nitrosoguanidine). Mutants can also be prepared by insertion, deletion or substitution of nucleotides, as well as, spontaneous mutation. The term Bacillus subtilis includes these mutants. The time of administration of the viable microorganism composition of Bacillus subtilis is particularly limited; however, in order to obtain proper colonization of the intestinal bacterial flora, it is preferred to administer the viable microorganism composition of Bacillus subtilis after administration of the above-described viable microorganism composition of lactic acid bacteria. Furthermore, in the case of a feeding method in which antibiotics, antibacterial and similar agents are not admired during a period between the stage of newborn chicks and the final stage, it is preferred to administer the viable microorganism composition of Bacillus. subtilis during that period. In the case of feeding of young chickens, for example, it can be administered during an optional period between the stage of newborn chicks and the final stage; however, when the drug-free feeding is performed only during the aging stage and the final stage, so that the drugs do not remain in the tender chicken body at the time of the tender chicken's shipment, the viable microorganism composition of the Bacillus subtilis can be administered during the breeding stage and final stage. The administration of the viable microorganism composition of Bacillus subtilis can be carried out by oral ingestion after addition to the food, drinking water or the like. For example, when the viable microorganism composition of Bacillus subtilis containing Bacillus subtilis C-31 02 is mixed with feed upon administration, it is preferred that the feed have a microorganism density of 1 to 1 to 08 viable microorganisms per gram in the form of spores and / or plant cells. The method of the present invention for admixing viable microorganism compositions for poultry, can be applied not only to chickens to be fed under heavy breeding in a large number of birds, but also to chickens to be fed under other conditions, and other domestic birds, such as ducks, geese, quails, wild ducks, ostriches and similar, as well as pet birds and the like. When compositions of viable microorganisms for poultry are administered in the method described above, an intestinal bacterial flora can be formed in the intestines, in which bacteria useful for the growth of poultry predominate., so that the poultry grow up in good health. Various effects can be obtained, such as, inhibition of growth of toxic bacteria, prevention of diarrhea, intensification of growth, improvement of the feed conversion ratios and similar, and the free feeding of drugs of chickens becomes possible. In addition, because the useful bacteria dominate the intestinal bacterial flora, the poultry grows in good health, so that delicious meat and eggs and the like can be obtained in the case of edible poultry, due to markedly improved qualities of the poultry. the products. In addition, meat, eggs and the like are safe and are not contaminated with toxic bacteria that poison food.

Examples Having generally described this invention, further understanding can be obtained by reference to certain specific examples, which are provided herein for purposes of illustration only, and are not intended to be limiting unless otherwise specified.

Production Example 1 A 4.5 g portion of skimmed milk was dissolved in 50 g of water, pasteurized at 1000 ° C for 10 minutes and then cooled to room temperature. A complete loop of Lactobacillus reuteri CP-720 was inoculated into the solution thus prepared and cultured statically for 24 hours at 37 ° C to obtain a first initiator (lactic acid bacteria: 1 x 1 08 viable microorganisms per gram). Then, 15 g of the first initiator was inoculated into 500 g of skimmed milk (solids content, 9% by weight), which had been pasteurized at 90 ° C and cultured for 20 hours at 37 ° C to obtain a second initiator. The second primer contained 2x1 08 viable microorganisms per gram. A medium prepared by dissolving 200 g of casein peptone, 200 g of yeast extract, 1 00 g of sodium citrate and 200 g of glucose in 20 kg of water, and adjusting the pH of the medium to 7.0 with sodium hydroxide solution. sodium 1 N, put in a jar fermenter, pasteurized at 95 ° C for 1 5 minutes and then cooled to room temperature. Subsequently, 3 parts by weight of the second initiator described above were inoculated in 1 00 parts by weight of the medium and cultured statically for 20 hours at 37 ° C. The culture broth thus obtained was centrifuged to recover the microorganisms, which were subsequently lyophilized using, as a dispersion medium, 1 kg of a solution containing 10% by weight of skimmed milk and 1% by weight of sodium glutamate, which had been pasteurized at 90 ° C in advance, thereby obtaining 146 g of viable microorganism powder from Lactobacillus reuteri CP-720. The viable microorganism powder contained 7.0x1 01 or viable microorganisms per gram.

Production example 2 A medium prepared by dissolving 200 g of beef peptone, 60 g of soy peptone, 1 00 g of yeast extract, 1 00 g of sodium acetate, 40 g of dipotassium phosphate, 60 g of diammonium citrate and 400 g of glucose in 20 kg of water and adjusting the pH of the medium to 7.0 with 1 N sodium hydroxide solution, put in a jug fermenter, pasteurized at 95 ° C for 1 5 minutes, and then cooled to room temperature. Subsequently, 500 g of a second Lactobacillus johnsonii CP-721 primer, which had been pre-cultured in advance by the same procedure as in Production Example 1, was inoculated into the medium to perform 1 8 hours of culture set at 35 ° C. ° C. The culture broth thus obtained was centrifuged to collect the microorganisms, which were subsequently lyophilized using, as a dispersion medium, a solution containing 10% by weight of skim milk and 1% by weight of sodium glutamate, which had it was pasteurized at 90 ° C in advance, thereby obtaining 141 g of viable microorganism powder from Lactobacillus johnsonii CP-721. The microorganism powder contained 6.5x1010 viable microorganisms per gram.

- Production Example 3 A second Lactobacillus reuteri CP-722 initiator prepared by the same procedure as in Production Example 1 was inoculated into a prepared medium and pasteurized in the same manner as described in Production Example 1, having the same composition and weight of components, and was cultured statically for 20 hours at 37 ° C.

The culture broth thus obtained was centrifuged to collect the cells, which were subsequently lyophilized using, as a dispersion medium, a solution containing 10% by weight of skimmed milk and 1% of sodium glutamate, which had been pasteurized at 95 ° C in advance, thereby obtaining 1 51 g of viable microorganism powder from Lactobacillus reuteri CP-722. The viable microorganism powder contained 6.8x1 01 or viable microorganisms per gram.

Production Example 4 The viable microorganism powders obtained in Production Examples 1 to 3 were combined in portions of one part and thoroughly mixed with 7 parts of dextrin to obtain a viable microorganism composition of lactic acid bacteria for poultry containing three strains of lactobacilli. The composition of viable microorganisms contained 2.0x1 010 viable microorganisms per gram.

Production Example 5 A medium prepared by dissolving 200 g of soy peptone, 1 g of dipotassium phosphate and 200 g of molasses in 1 g of water and adjusting the pH of the medium to 7.5 with 1 N sodium hydroxide solution. , it was placed in a jar fermenter, pasteurized at 95 ° C for 60 m inutes and then cooled to 37 ° C. Subsequently, 1 00 g of a culture broth of Bacillus subtilis C-31 02, which had been pre-cultured in advance, was inoculated into the medium and agitated aerobically for 40 hours at 37 ° C. The culture broth thus obtained was centrifuged to recover the microorganisms, which were subsequently mixed with the same weight of skimmed milk and lyophilized under vacuum, thereby obtaining 750 g of the viable microorganism composition of Bacillus subtilis for poultry. The viable microorganism composition of Bacillus subtilis for poultry contained 1.2 x 10 010 viable microorganisms per gram.

Example 1 Field effect test I (test without drugs in and after the breeding stage) The test is carried out using Chunky (commercial name, commercial strain). Chickens (0 days after hatching) in a carrier cage of chickens were sprayed with 12 g of the composition of viable microorganisms of lactic acid bacteria for poultry prepared in Production Example 4 (total number of lactic acid: 2.0x1 01 0 viable microorganisms per gram), which have been uniformly suspended in 4 kg of pure water. As the provision food, "Chick Prestarter Feed (H iña Ezuke)" was used for initial feeding use (containing antibiotics) and "Starting AT (Zenki AT)" for use of start stage (containing antibiotics), in the stage feeding start (from 0 to 5 days after hatching) and starter stage (from 5 to 21 days after hatching), respectively. "Brogoal A" was used for final use (without antibiotics), which had been mixed with 9x1 05 cells / g of the viable microorganism composition of Bacillus subtilis for poultry, prepared in Production Example 5, during the period from the aging stage (from 21 days to 43 days after hatching) to the final stage (from 43 days to the day of shipment) (all these commercial food items are manufactured by Chubu Shiryo). Another feed management was carried out according to the method of feeding chickens conventionally carried out in poultry farms. The results of the test are shown in Table 2.

Comparative Example 1 Feeding was carried out in the same manner as in Example 1, except that the administration of the viable microorganism composition of lactic acid bacteria was not performed, a "Grower AT (KOHKI AT)" was used for the use of the aging stage (containing antibiotics) (manufactured by Chubu Shiryo, Co.), as the food in the aging stage instead of the "Brogoal A" for final use, and the viable microorganism composition of Bacillus subtilis is not provided throughout the complete stages. The results are shown in Table 2. In this connection, the "Grower AT (Kohki AT)" is a food item having the same composition as that of "Brogoal A", except for antibiotics.

Table 2 Breeding proportion: (the number of birds on board / the number of chickens in the set) x 1 00 Feed conversion ratio: (total amount of food eaten during feeding (g) / total body weight gain (g)) 1 00 Production result: (average body weight x breeding ratio / feed conversion ratio x age in days when they were shipped) x 1 00 The value of the average body weight gain in Example 1 was less than that of Comparative Example 1 by a factor of about 1%, which was assumed to be because of the quality of the chickens tested. This, the chicken body weight was standard (41 .9 g / chicken) in the shared Example 1, but was close to that of the smaller chickens (38.4 g / chicken) in Example 1, so that its growth became lower. Because other results of Example 1 were superior to those of Comparative Example 1, productivity was improved.

Example 2 Field effect test I I (test without drugs in and after the start stage) The step was carried out using Chunky (trade name, commercial strain). Chickens (0 days after hatching) in a carrier cage of chickens were sprayed with 12 g of the viable microorganism composition of lactic acid bacteria for poultry prepared in Production Example 4 (total number of bacteria of lactic acid: 2.0x1 01 0 viable microorganisms per gram), which had been uniformly suspended in 4 kg of pure water. As the provision food, the "Chick Prestarte Feed" was used for initial feed use in the initial feeding stage (from 0 to 5 days after hatching). "Starting AT (Zenki AT)" was used for the start stage from which the antibiotics had been removed and the "Brogoal A" for final use, which had been mixed with 9x1 05 visible microorganisms per gram of the composition of viable microorganisms of Bacillus subtilis for poultry prepared in Production Example 5, in the initiating stage (from 5 to 21 days after hatching) and during the period from the breeding stage (from 21 days to 43 days) after the hatching) until the final stage (from 43 days until the day of shipment), respectively. Another feed management was carried out according to the method of feeding chickens conventionally carried out in poultry farms. The results of the test are shown in Table 3.

Comparative Example 2 Feeding was performed in the same manner as described in Example 2, except that "Starting AT (Zenki AT)" was used for the initial stage use and "Grower AT (Kohnki AT)" for use by the breeding stage, in the initial stage and breeding stage, respectively, and the viable microorganism composition of Bacillus subtilis was not provided throughout the complete stages. The results are shown in Table 3.

Table 3 Because drug-free food was used in and after the initiation step in Example 2, while food containing medicaments was used in Comparative Example 2, the body weight gain in the previous case was slightly lower with a value of 51.6 g / day, but the breeding rate was 97%, which was excellent and the production result was 244, showing results of economic production.

Example 3 Field effect test l l l (test without drugs) The test was performed using Chunki (trade name, commercial strain). Chickens (0 days after hatching) in a carrier cage of chickens were sprayed with 40 g of the viable microorganism composition of lactic acid bacteria for poultry prepared in Production Example 4 (total number of bacteria of lactic acid: 2.0x1 01 0 viable microorganisms per gram), which had been uniformly suspended in 4 kg of pure water. As the provision food, the "Starting AT (Zenki AT)" for use of the start stage from which the antibiotics had been removed and the "Brogoal A" for final use, which had been mixed with 9x1 05 m icroorganisms viable per gram of the viable microorganism composition of Bacillus subtilis for poultry prepared in Production Example 5, were used during the period from the initial feeding stage to the starting stage (from 0 to 21 days after hatching) and during the period from the breeding stage to the final stage (from 21 days to the day of boarding), respectively. Another feed management was performed according to the chicken feeding method conventionally carried out in poultry farms. The results of the test are shown in Table 4.

Comparative Example 3 The feeding was performed in the same manner as described in Example 3, except that the "Starting AT (Zenki AT)" for the use of the start stage, the "Grower AT (Kohki AT)" for stage use. and the "Brogoal A" for final use, respectively were used as the food during the period from the initial feeding stage to the start stage, in the breeding stage (from 21 to 43 days after hatching) ) and in the final stage (from 43 days on the day of boarding), respectively, and the viable microorganism composition of Bacillus subtilis was not provided throughout the complete stages. The results are shown in Table 4.

Table 4 The test of Example 3 was not performed at the same time as the test of Comparative Example 3 but after the end of the feed of Comparative Example 3, and, because a cold wave hit the field during the feeding period of Example 3, the final growth and breeding ratio in Comparative Example 3 were influenced by the cold wave. However, although feeding was performed without administering antibiotics and antibacterial agents over the entire period, the production results of Example 3 were identical to those of Comparative Example 3. In addition, the comparative tests corresponding to Examples 1 to 3, in which the feeding without drugs and without administering the composition of viable microorganisms of lactic acid bacteria for poultry and the composition of viable microorganism of Bacillus subtilis for poultry, were not carried out, due to the fact that obtained normal growth under such feeding conditions.

EXAMPLE 4 In Example 3 and Comparative Example 3, fresh droppings of chickens were collected on the day before the shipment and, using droppings of three birds as a sample, the group of coliforms and bacteria of the genus Salmonella were measured in the droppings. The results are shown in Table 5. In this case, the serotype of each bacterium of the genus Salmonella was O7.

Table 5 * p < 0.05 ** p < 0.01 Compared to Comparative Example 3, the group of coliforms decreased significantly in Example 3, and the detection rate of bacteria of the genus Salmonella was also significantly reduced.

Example 5 A comparison was made on the acceptability of the meat of chickens bred in Example 3 and Comparative Example 3. Because a precise sensory test can not be performed when the meat itself is subjected to the test due to various treatment factors, whole chicken soup was prepared and subjected to the sensory test. The intestines III (remaining edible part after slaughter and bleeding and subsequent removal of feathers, head, legs and organs) were used in the test as the whole chicken, and one part by weight of the sample was mixed with 4 parts by weight of water and 1.8% salt and boiled for 2 hours. After filtering the mixture through blanched cotton, the filtrate (soup) was prepared three times the weight of the chicken tested to adjust the concentration of the soup. The soup samples thus obtained were subjected to the sensory test, in which 38 panelists were allowed to select the most delicious sample. The results are shown in Table 6.

Table 6 p < 0.01 Compared to the chicken of Example 3, chicken 3 was selected for its acceptability by a considerably large number of panelists, thus confirming the delicious nature of the chicken produced through drug-free feeding. In this connection, in a specified flavor evaluation conducted at the same time, good evaluations were obtained for the chicken soup of Example 3, statistically significant with a level of significance of 1%, in each of (1) preference, ( 2) delight in flavor and (3) adequate flavor.

Reference Example 1 Completely drug-free test only by viable microorganism composition of Bacillus subtilis for poultry The test was performed using Chunky poultry in a district where young chickens are fed in large numbers. As the provision food, "Broiler Gold Starting Crumble" (from Broiler Gold Zenki Crumble), from which the antibiotics had been removed, and "Broiler finisher (Broiler Shiage)" (without containing antibiotics and the like) (both manufactured by Shikoku Haigo) were used. Shiryo, Co.), each of which had been mixed with 1 x1 06 viable microorganisms per gram of the viable microorganism composition of Bacillus subtilis for poultry prepared in Production Example 5 and enriched with vitamins and minerals, during the period from the initial feeding stage to the starting stage (from 0 to 21 days after hatching) and during the period from the breeding stage to the final stage (from 21 days after hatching until the day of em barque), respectively. Another handling of the feed was carried out according to the method of feeding chickens conventionally carried out in poultry farms. The results of the test are shown in Table 7.

Comparative Example 4 Feeding was carried out in the same manner as in Reference Example 1, except that "Broiler Gold Starting Crumble (Broiler Gold Zenki Crumble)" (containing antibiotics), "Broiler S" (containing antibiotics) and "Broiler Gold" were used. Broiler Finisher (Broiler Shiage) "(each of which is manufactured by Khikoku Haigo Shiryo, Co.), as the feed during the period from the initial feeding stage to the start stage, in the breeding stage (from 21 up to 32 days after hatching) and in the final stage (from 32 days after hatching until the day of shipment), respectively, and the viable microorganism composition of Bacillus subtilis was not provided. The results are shown in Table 7.

Table 7 A higher breeding ratio was obtained in Reference Example 1 than in Comparative Example 4, but the body weight gain was delayed in the feeding start stage and the delay left a trace at the end, so that the weight gain body was low throughout the feeding period as a result of it, and was not an acceptable commercial level. In addition, a drug-free breeding trial was not performed by providing only the viable microorganism composition of lactic acid bacteria for poultry without administering the viable microorganism composition of Bacillus subtilis for poultry, because such a test can be performed only in a place where environmental pollution does not exist, such as an experimental facility under complete sanitary conditions but not in the field of intensive poultry farms.

INDUSTRIAL APPLICABILITY Because the administration of a viable microorganism composition for poultry, comprising specific viable lactic acid bacteria, is carried out at a specified stage in combination with the administration of a viable microorganism composition for poultry comprising Bacillus subtilis, the method of the present invention for the administration of viable microorganism combinations for poultry can exert various effects, such as, inhibition of the growth of toxic bacteria, prevention of diarrhea, intensification of growth, improvement of the feed conversion ratios and the like, and also makes it possible to feed free medicines of chickens with high productivity and, in particular, in the case of edible chickens, the production of delicious and safe meat and eggs with high productivity. Accordingly, the method of the present invention for the administration of viable microorganism compositions for poultry is useful for feeding edible birds, such as, chickens, particularly, young chickens, and the like. Obviously, numerous modifications and variations of the present invention are possible in light of the above teachings. Accordingly, it will be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described herein. The priority document of the present application, Japanese patent application no. Hei 9-145372, filed June 3, 1992, is incorporated herein by reference.

Claims (26)

1. REIVI N DI CATIONS 1 . A method for administering viable microorganism compositions to poultry, comprising: administering a first composition to a poultry, comprising viable bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii, in the stage of newborn chicks; and administering a second composition to said poultry, comprising a viable microorganism belonging to Bacillus subtilis.
2. 2. The method according to claim 1, wherein said first composition is administered within 4 days after said poultry hatched.
3. 3. The method according to claim 1, wherein said first composition is administered within 2 days after said poultry hatched. The method according to claim 1, wherein said Lactobacillus reuteri is at least one member selected from the group consisting of (a) Lactobacillus reuteri CP-720, Deposit No. FERM BP-6332, and a mutant thereof, (b) Lactobacillus reuteri CP-722, Deposit No. FERM BP-6334, and a mutant thereof. The method according to claim 1, wherein said Lactobacillus johnsonii is Lactobacillus johnsonii CP-721, Deposit No. FERM BP-6333, or a mutant thereof. 6. The method according to claim 1, wherein said first composition further comprises a carrier or solvent. 7. The method according to claim 1, wherein said first composition further comprises a feed for poultry. The method according to claim 1, wherein said first composition is administered orally. The method according to claim 1, wherein said first composition is administered by atomization. The method according to claim 1, wherein said first composition comprises from 10d to 1010 of said bacteria per gram. eleven . The method according to claim 1, wherein 1 x 10 3 to 1 x 1 0 0 of said bacteria are administered to said poultry. 12. The method according to claim 1, wherein said first composition is administered only once or twice. The method according to claim 1, wherein said Bacillus subtilis is Bacillus subtilis C-31 02, Deposit No. FERM BP-1 096, or a mutant thereof. The method according to claim 1, wherein said second composition further comprises a carrier or diluent. 15. The method according to claim 1, wherein said second com- position further comprises a feed for poultry. The method according to claim 1, wherein said second composition is administered orally. The method according to claim 1, wherein said second composition comprises from 105 to 1000 of said viable Bacillus subtilis per gram. 18. The method according to claim 1, wherein said second composition is administered after the administration of said first composition. 9. The method according to claim 1, wherein said poultry is a chicken, a duck, a goose, a quail, a wild duck, an ostrich or a pet bird. The method according to claim 4, wherein said Lactobacillus johnsonii is Lactobacillus johnsonii CP-721, Deposit No. FERM BP-6333, or a mutant thereof, and said Bacillus subtilis is Bacillus subtilis C-31 02 , Deposit No. FERM BP-1 096, or a mutant thereof. twenty-one . A method for administering viable microorganisms to poultry, comprising: administering a first composition to a poultry comprising viable bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii, in the stage of newborn chicks. The method according to claim 21, wherein said Lactobacillus reuteri is at least one member selected from the group consisting of (a) lactobacillus reuteri CP-720, Deposit No. FERM BP-6332, and a mutant thereof, (b) Lactobacillus reuteri CP-722, Deposit No. FERM BP-6334, and a mutant thereof. 23. The method according to claim 21, wherein said Lactobacillus johnsonii is Lactobacillus johnsonii CP-721, Deposit No. FERM BP-6333, or a mutant thereof. 24. A poultry in the newborn pigeon stage, prepared by the method of claim 21. 25. A poultry in the newborn pigeon stage, prepared by the method of claim 22. 26. A poultry in the newborn pigeon stage, prepared by the method of claim 23. SUMMARY A method for administering viable microorganism compositions for poultry, comprising: administering a viable microorganism composition comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii in the stage of newborn chicks; and administering a viable microorganism composition, which comprises a viable microorganism belonging to Bacillus subtilis.