JP2001149023A - Bioactive agent and animal feed additive using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To improve the appetite of densely bred livestock, enhance the digestion rate of roughage, and promote the digestive absorption of nutrients. SOLUTION: This bioactive agent for improving a fattening rate, meat quality and so on is obtained by together culturing lactobacillus and yeast in a specific combination ratio, and has effects such as the saving of feed costs. The bioactive agent further reduces the bad smell of the livestock itself as a main cause for the generation of a characteristic bad smell in a stock-raising process.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel bioactivity which contributes to the improvement of growth and quality and stimulates immunity by administering or spraying a culture of cultivated and adapted bacteria to animals and plants. Agent. The culture described in the present text mainly refers to the culture solution itself and only the cells, but the culture filtrate from which bacteria have been removed also has physiological activity and is therefore included in the category of culture.

[0002]

2. Description of the Related Art In the livestock industry, in order to improve the growth rate of livestock, it has become common to feed livestock containing high fat and high carbohydrates to the animals to suppress the energy consumed by exercise and carry out overcrowding. ing.

[0003] However, in livestock placed in such an environment, the constitution of the livestock is acidified and various adverse effects due to so-called acidosis such as a decrease in appetite and deterioration of meat quality are likely to occur. However, in such livestock, the growth rate is reduced, and further, in the obtained meat or the like, the concentration of peroxide is increased, the freshness holding power is reduced, and the efficiency is reduced. There was a problem that good breeding and good quality meat could not be obtained. Further, in the conventional breeding method, a given feed may be discharged in an undigested state, and therefore, there is a problem that the livestock barn has a bad odor and this causes pollution.

[0004] In addition, similar harmful effects exist in fish cultivation, and furthermore, pathogenic bacteria and toxins produced due to the spoilage of the residual feed that has settled on the bottom cause mass death and economic loss. There is something immeasurable.

[0005] The same applies to the cultivation of insects, earthworms and the like, albeit of a large or small scale.

[0006] Turning to the field of agriculture, especially in large-scale intensive cultivation, large amounts of chemical fertilizers and pesticides have been used. At present, there are environmental problems such as pollution of groundwater and rivers, and also health problems of agricultural workers.

[0007]

The problem to be solved by the present invention has been conventionally solved by administering or spraying the present bioactive agent while substantially following conventional intensive breeding, cultivation or cultivation methods. To provide an activator intended to promote healthy growth and produce high-quality products that include high yield and color luster while ensuring high yield while reducing adverse effects. It is in.

[0008]

Means for Solving the Problems As a result of repeated investigations to achieve the above object, the present inventor has found that the reincarnation of nature is growth and collapse, and biological substances are no exception. The most important purification system operates smoothly and smoothly, and the removal of various negative factors that inhibit the ability of the organism itself, including the absorption and synthesis of the organism, removes the organism's original form, and therefore has various problems. Most of them were expected to be resolved, and under these expectations, we worked diligently. Finally, a system that condenses the purification of the natural world, that is, an activated sludge method that skillfully utilizes useful microorganisms for the purpose of environmental restoration, was applied, and this system was also applied to biological purification, and the present invention was achieved. Achieved.

That is, according to a first aspect of the present invention, there is provided a bioactive agent comprising a culture solution obtained by co-culturing lactic acid bacteria and yeast, wherein the culture solution comprises lactic acid bacteria and yeast, respectively. And the ratio of the number of lactic acid bacteria to the number of yeast cells per unit amount of the culture solution is 100: 10 to 10:10.
A bioactive agent characterized by being in the range of 0:30.

[0010] The present invention according to a second aspect is a bioactive agent obtained by mixing culture solutions obtained by separately culturing lactic acid bacteria and yeast cells, wherein the mixed culture solution contains lactic acid bacteria and yeast cells. The ratio of the number of lactic acid bacteria to the number of yeast cells per unit amount of the mixed culture solution containing the cells is 100: 10 to 1
It is a bioactive agent characterized by being in the range of 00:30.

According to a third aspect of the present invention, there is provided the culture medium of the first invention or the mixed culture medium of the second invention, wherein the culture medium is further selected from the group consisting of inorganic vegetative bacteria, photosynthetic bacteria, and bacteria of the genus Bacillus. A bioactive agent to which at least one or more bacterial cells are added, wherein the bacterial cells are contained in a range of 10 to 50 with respect to 100 lactic acid bacteria per unit amount of the bioactive agent. It is a characteristic bioactive agent.

According to a fourth aspect of the present invention, the inorganic vegetative bacterium according to the third aspect is characterized in that the bacterium is Nitrobac (Nitrobac).
ter), Nitrosomonas
s), Thiobacillus, and Thiobacterium
A bioactive agent characterized by being a bacterium selected from the group of:

According to a fifth aspect of the present invention, there is provided the method according to the first or second aspect, wherein the culture solution or the mixed culture solution further comprises Bacillus subtilis.
lis) and Bacillus mesentericus (Bacill)
The present invention is a bioactive agent characterized by being prepared by adding and mixing cells of L. mesentericus in the range of 20 to 40 and 10 to 20 with respect to 100 lactic acid bacteria, respectively.

According to a sixth aspect of the present invention, there is provided a bioactive agent according to the first to fifth aspects, wherein the lactic acid bacterium contains at least one bacillus lactic acid bacterium and at least one cocci lactic acid bacterium.

According to a seventh aspect of the present invention, there is provided a bioactive agent comprising a culture filtrate obtained by removing cells from a culture solution or a mixed culture solution according to the first to sixth aspects.

According to an eighth aspect of the present invention, there is provided an animal feed additive obtained by shaping the bioactive agent according to any one of the first to seventh aspects with an excipient. .

According to a ninth aspect of the present invention, there is provided the animal feed additive according to the eighth aspect, wherein the animal feed additive is 10 to 30% by weight based on the feed.
Animal feed characterized by being added.

The present inventor has searched from the viewpoint of what bacteria are most suitable for biopurification. As a result, the pathogen is non-pathogenic, and can be used in the intestine, oral cavity, and other living organisms to obtain grass, leaves, agricultural products, fruits, fermented foods, soil, and the like. Lactic acid bacteria that have a very wide range of nature in the living sphere, down to the sewage, are compatible with the lactic acid bacteria, and are abundant in plants, wild yeasts, and cultured yeasts that have been kneaded in the fermentation industry. Was selected, these bacteria were collected as much as possible, and the mixed culture was continued for about one year.

Further, to these bacteria, inorganic vegetative bacteria (bacteria which exhibit plant metabolism and are non-pathogenic) and bacteria of the genus Bacillus (saccharifying bacteria, proteose-producing bacteria) which are active in activated sludge are added. Symbiotic subculture was repeated.

During the passage, there are many bacteria that are lost due to growth competition and disappear depending on the species and strains, and there are many yeasts that are aggregated and dropped off by lactic acid bacteria. Furthermore, they succeeded in creating a group of useful microorganisms in which only bacterial species and strains that established a symbiotic relationship inhabit.

Furthermore, regardless of the type of bacteria and the number of bacteria finally symbiotic in these useful microorganism groups, regardless of the number of bacteria in the seed starter, a certain co-cultivation period (for example, a specific temperature) After about one week to several weeks under the same conditions), it is found that the stability becomes almost constant.
Therefore, the symbiotic culture obtained after the symbiotic culture for about one year and the symbiotic culture obtained by the symbiotic culture under a specific temperature or the like are substantially similar in terms of the effects and effects of the present invention. They have found that they have a tendency and have achieved the present invention.

When cultures of these useful microorganisms are mixed with feed and fed to experimental animals such as mice, rats, rabbits, etc., they have a growth promoting effect, have a good appearance such as color and luster, and have a good appearance. Confirmed that the resistance to pathogens such as cattle, pigs, chickens, and other livestock, poultry, and various fish such as carp, eel, sweetfish, hamachi, and Thailand, as well as beetles, stag beetles, Insects such as insects or their larvae, earthworms and mosquitoes were administered to various animals, and good results were obtained.

Similarly, when plants are first germinated and sprayed on radish (Japanese radish) and Komatsuna several days later in an appropriate amount, the plants not only grow faster but also grow faster than non-sprayed plants. And the growth of leaves improve
It can be confirmed that a large amount of good quality is harvested, and it is used for various plants, from vegetables such as strawberries, tomatoes, and spinach, to fruit trees such as tangerines, fruits, lawns and houseplants. I got a good result.

Incidentally, the following effects were also obtained as a secondary effect. For example, in animal breeding, it is not an exaggeration to say that the treatment of excrement is the biggest issue. Some places are frowned upon as the greatest offensive odors, but the administration of this drug has already reduced the odor at the time of excretion. Fermentation of the excrement progresses with the passage of time, and the odor further decreases, so that there is almost no pollution problem.

It was also found that by spraying foliarly on plants, the useful microorganisms contained in the present agent adhere to and survive on the plants, which hinders the chances of infection with mold and plant pathogenic bacteria. For example, downy mildew, rust, mildew, powdery mildew, etc. can be defended with considerable probability.

For the culture in this case, 100 ml of yeast extract was added to 1000 ml of neutral pH water as a medium.
g, CGF (chlorella gross factor) 50 mg,
Trehalose 50 mg, lactose 25 mg, KH ₂ PO
₄ 15mg, MgSO ₄ · 7H ₂ O to 15 mg, Na
10 mg of Cl was mixed to form a medium, and cultured at 28 ° C. to 32 ° C. for 10 to 15 days.

As the inoculum of the mixed bacterium, a 5-liter glass triangular corben was used, into which 5 liters of the above-mentioned medium was poured, and subjected to high-pressure sterilization at 120 ° C. for 15 minutes. Was suspended in 1 ml of sterile physiological saline, and the suspension was mixed, inoculated into the above-mentioned medium, and cultured according to the above-mentioned culture method to obtain a seed of a mixed bacterium.

In the present invention, the coexistence of lactic acid bacteria and yeast is essential. The reason is that secretions of lactic acid bacteria promote the growth of yeast, and conversely, secretions of yeast promote the growth of lactic acid bacteria. This is because, in the present invention, since lactic acid bacteria and yeast are co-cultured, compared to when each bacterium is cultured alone, breeding is promoted and the culture efficiency can be increased. .

Then, when the main culture solution was mixed with and fed to experimental animals such as mice, rats, and rabbits and fed, it was found that it had a growth promoting effect. In addition, the present invention has been completed by finding that animals such as pigs, cows, chickens and other livestock and poultry have an effect of promoting the growth of animals when mixed with feed and fed.

The animals to be used in the present invention include livestock,
Animals such as poultry, fish, etc.
Domestic animals such as chickens, deer, rabbits and goats, monkeys, mice, rats, experimental animals such as guinea pigs, various fish such as carp, eel, sweetfish, coho salmon, Thailand, hamachi, flounder,
Particularly preferably, the period of remarkable growth from the young body is targeted.

[0031]

DESCRIPTION OF THE PREFERRED EMBODIMENTS As described above, the bacterial species and the bacterial ratio of a group of useful microorganisms that have established a symbiotic relationship after growing and acclimating for about one year have been investigated and measured. Lactobacillus casei (Lactob)
acillus casei), Lactobacillus acidophilus (Lactobacillus acido)
philus), Lactobacillus plantarum (La)
Cactobacillus plantarum, Lactobacillus caucasians (Lactobacillus)
scaucasicus), Lactobacillus lactis, Lactobacillus thermophilus (Lactobacill)
lac thermophilus is established and symbiotic, and as lactic acid bacteria, Streptococcus lactis (Streptococcus lactis), Streptococcus cremoris (Streptococ)
cus cremoris, Streptococcus thermophilus (Streptococcus ther)
mophilus), Streptococcus diacetiactis (Streptococcus diacet)
iactis), Streptococcus bulgaricus (Streptococcus bulgaricu)
s), Enterococcus feucharis (Enteroc)
occus faecalis), Leuconostoc
Cremoris (Leuconostoc cremori)
s), Leuconostoc Mesenteroides (Leuc)
onostoc mesenteroides), Pediococcus halophilus (Pediococcus)
halophilus) had established colonization.

[0032] Examples of yeasts include Saccharomyces cerev.
siae), Saccharomyces fragilis (Sacc)
haromyces fragilis, Saccharomyces ellipsoidis
ellipsoidus), Saccharomyces awamori
i), Saccharomyces carlsbergensis (Sac
charomyces carlsbergensi
s), Saccharomyces luxii (Saccharom)
yces rouxii), Saccharomyces florentims
timus), Saccharomyces salmon (Sacchar)
omycessake), Candida arborea (Ca
ndida arborea, Candida utilis, Candida tropicalis, and Rhodotorula were established and symbiotic.

As the inorganic vegetative bacterium, a digestive bacterium such as the genus Nitrobacter or Nitrosomonas, a sulfur bacterium such as the bacterium Thiobacillus or Tiobacterium, or a photosynthetic bacterium (Achiorho daceae spp.) Can be used.
However, other bacteria belonging to the genus Bacillus include Bacillus subtilis and B. mesentericus.
us) lived together.

The ratio of the bacterial flora and the number of bacteria once constructed does not fluctuate greatly even after repeated passage under the same conditions, as in the case of the intestinal flora.
In the range of 100: 10 to 100: 30, lactic acid bacteria:
Yeast: inorganic vegetative bacteria and Bacillus genus = 100: 10:
The range was 10 to 100: 30: 50, and it was found that the use of the bacteria with a ratio of the number of bacteria within this range was more effective.

Even when cultivation is performed under other conditions (for example, when the culture medium, temperature, and cultivation time are changed), if the bacterium ratio is adjusted to the above-mentioned ratio, the efficacy as the present agent is not significantly different. Absent.

It is to be noted that, even if the above-mentioned growth acclimatization method is not adopted, the same bacterial species are cultured and the number of lactic acid bacteria and the number of yeasts are determined.
Or, the ratio of the number of lactic acid bacteria and the number of yeasts, and the number of bacteria such as inorganic vegetative bacteria, etc., within the above range, although the titer is inferior, shows a significant difference compared to the case of not using This was also confirmed.

It is also clear that the minimum unit as a useful microorganism group is a necessary condition that it is a culture composed of at least one kind of bacterium lactic acid bacterium, one kind of bacillus lactic acid bacterium, and one kind of yeast. Was. These lactic acid bacteria, yeast, and inorganic vegetative bacteria, etc., are each separately subjected to slant culture, and the bacteria cultured here are added to sterile physiological saline to form a bacterial suspension. Mix and expand in coexistence.

The total number of these bacteria coexisting in the co-culture solution thus obtained is preferably 10 7 or more per milliliter of the culture solution, and the pH is also 4.0.
It is preferably maintained at about 5.5.

[0039] In this case, the liquid medium is desirably low in nutrients as much as possible, and it is important to train under low nutrients.

[0040] The reason is that, in a severe environment in the natural world, if the bacterium cannot survive the growth competition with other fungi and grow, it cannot survive at all.

As a blending example, it is preferable to add 0.5 to 0.8 parts by weight of glycine to 1 part by weight of peptone. In this case, as an example, lactose 15 to 20 g, glucose 30 to 40 g, salt 2 to 5 g, peptone 20 to 30 g, glycine 15 to 20 per 1000 liters of water.
g can be added to obtain the liquid medium of the present invention. It is also possible to add molasses or fruit water instead of glucose.

Further, the bacteria are separately cultured and grown and acclimated, respectively, and the culture solution is centrifuged, and the bacteria are mixed in the above ratio or freeze-dried according to a conventional method. The bacteria may be mixed and used.

When co-culture is carried out, yeast functions to convert pyruvate into glucose by using glucose as a substrate and primary metabolism of glycolysis, and pyruvate is converted into aerobic acetic acid bacteria and microaerobic lactic acid bacteria. It becomes a substrate for anaerobic butyric acid bacteria, and in the secondary metabolism, it produces organizing active substances such as hormones, vitamins, nucleic acids, etc., and these lactic acid bacteria, yeasts, etc. maintain and complement each other and grow together. can do.

The additive according to the present invention may be in any form, for example, liquid, powder, granule, tablet and the like. When the co-culture medium is used without drying as an additive, the additive may be in a suspended state or a paste form.

Specifically, cyclodextrin powder was added in an amount of about 0.2 to 0.7 with respect to 1 part by weight of the obtained co-culture solution.
Add parts by weight and stir at high speed to form a paste. If the amount of the cyclodextrin powder is less than 0.2 parts by weight, the effect of inclusion is insufficient, and if it exceeds 0.7 parts by weight, the addition becomes excessive. When the paste obtained here is spray-dried using a spray dryer, the target powder is obtained.

Further, a water-absorbing additive such as sawdust, zeolite, or foamed concrete is directly mixed with the co-culture solution to adjust the water content, or the water content is removed to obtain a solid dry bacterial cell to obtain a powder or granule. Alternatively, a method of using the composition as a tablet may be used. Any known excipients can be used as the excipient when the co-culture solution is solid. For example, those soluble in water include glucose, lactose, cyclodextrin, saccharides such as oligosaccharides or soluble starch, albumin,
Proteins or salts such as casein are used, and those insoluble in water include starch, bran, sawdust, bacas and other starches, natural materials mainly containing cellulose, chitin, gelatin, etc., calcium carbonate, calcium sulfate. And inorganic materials such as magnesium carbonate, activated carbon, white sand, diatomaceous earth, and glass.

The additives used in the present invention may be added alone to the feed, but may be mixed with casein powder, skim milk powder, molasses, etc. in order to improve palatability (preference such as biting). .

For plants, various commercially available inorganic and organic fertilizers and soils such as humus can be used as excipients.

In the additive thus prepared,
The co-existing cells of the present invention may be prepared at an appropriate content. For example, 1 × 10 5 cells / g per 1 g of animal feed additive
It may be prepared as about 1 × 10 12 cells / g of viable bacteria. The method of using the animal feed additive of the present invention,
Either method may be used. For example, it may be administered by mixing it in a feed, or may be administered as a liquid suspended in drinking water or milk.

[0050]

Next, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

1. Examples of animal feed additives

[0052]

Example 1 Lactobacillus casei, Lactobacillus acidophilus (Lactobacillus) were used as lactobacillus inoculum.
us acidophilus, Lactobacillus plant
arum) and Lactococcus inoculum as Streptococcus lactis (Streptococcus lact)
is), Streptococcus cremoris (Strep)
tococcus cremoris, Streptococcus thermophilus (Streptococcus)
thermophilus), Streptococcus
Diacetiactis (Streptococcus d
iacentiactis), Leuconostoc mesentere
roides), and Saccharomyces cerevisiae (Saccharomycesc) as the yeast inoculum.
erevisiae).

Each of these inoculum is individually cultured by agar culture to obtain a seed starter and a mother starter. Sterile water or sterile saline is added to each obtained colony to form a cell suspension. These individually prepared suspensions are mixed and expanded in the presence of each bacteria (usually primary expansion and secondary expansion)
I do.

The expansion culture was performed with 0.3% peptone and 0.1% peptone.
Lactobacillus casei about 1.5 ml, Lactobacillus acidophilus about 100 ml as a bacterium-type lactic acid seed in 100 ml of a substrate containing 5% yeast extract, 2% skim milk powder, and 0.5% calcium carbonate 0.0
5 ml, Lactobacillus plantarum approx.
About 15 ml, about 0.3 ml of Streptococcus lactis, about 0.1 ml of Streptococcus cremoris, about 0.1 ml of Streptococcus thermophilus,
About 0.05 ml of Streptococcus diacetiactis, about 0.1 ml of Leuconostoc mesenteroides, and about 0.2 ml of yeast as a Saccharomyces cerevisiae, 28
Incubate at 24 ° C. for 24 hours to obtain a seed starter. Next, it was added to 3 liters of the same substrate,
Incubate for a time to obtain a mother starter. Next, water 100
20 g of lactose, 30 g of glucose, 5 g of salt, 20 g of peptone and 15 g of glycine were added to 0 liter to prepare a liquid medium, and 10 liters of the mother starter was added to 1 ton of the liquid medium at 30 ° C. 1 week ~
After performing the main culture for 2 weeks, the ratio of viable bacteria in the culture solution was examined. As a result, the ratio of lactic acid bacteria: yeast was about 100: 15.

After adding 0.4 parts by weight of cyclodextrin to 1 part by weight of the obtained culture solution, the mixture was stirred at a high speed and spray-dried using a spray dryer to obtain a powder containing the components of the culture solution. (Hereinafter referred to as Formulation 1).

[0056]

Example 2 A mother starter was prepared in the same manner as in Example 1, and then lactose 25 was added to 1000 liters.
g, glucose 35 g, salt 5 g, peptone 30 g, and glycine 20 g were added to prepare a liquid medium. To 1 ton of the liquid medium, 10 liters of the mother starter was added, and the mixture was added at 30 ° C. for 1 to 2 weeks. After culturing, the ratio of viable bacteria in the culture solution was examined.
00:25. A powder containing the components of the co-culture solution was obtained in the same manner as in Example 1 (hereinafter, referred to as Formulation 2).

[0057]

Example 3 In addition to yeast, Bacillus subtilis (Bacillus subt.)
ilis) and Bacillus mesentericus (Bacil)
mesentericus) in the same manner as in Example 2, and the ratio of viable bacteria in the culture solution was examined. As a result, the ratio of lactic acid bacteria: yeast: Bacillus subtilis: Bacillus mesentericus was about 100: 10: 30: 15. Met. Next, in the same manner as in Example 1, a powder containing the components of the co-culture solution was obtained (hereinafter, referred to as formulation 3).

2. Example of breeding

[0059]

Example 4 A breeding test was performed using rats. The rats were bred under the conditions of breeding temperature of 22 ° C. (± 0.5 ° C.), free water supply, and free feeding. After pre-breeding for one week, rats without abnormal appearance and behavior were confirmed (SD rats: CLEA Japan, Inc.). , Male, 200 g at the time of introduction), and 13 test plots were set with 10 test gauges per gauge as one test plot. Feed and drinking water were freely available and fed ad libitum. The feed fed was a rat breeding feed, mixed with the additives prepared by the methods of Examples 1 to 3, and processed into pellets. Thirteen types of test feed pellets were prepared in total, and feed 1 had a weight ratio of additives of about 5% to feed, feed 2 had about 10%, feed 3 had about 30%, and feed 4 had about 40%. , And an additive-free feed was prepared as a feed 5 as a control. The test period was 3 weeks, and the weight of the rats was measured every week.

As a result of the above experiment, there was no death in any of the administration groups, and in the observation of the symptoms, feeds 2, 3 and 4 were observed.
In the test group to which each of the feeds was fed, no abnormal changes such as illness and diarrhea were observed even when compared with the control group to which no additive was added. No significant difference was found between the two. Table 1 shows the changes in feed weight and the rate of increase.

[0061]

[Table 1]

As shown in Table 1, the increase was observed in the 2nd and 3rd sections as compared with the control section, and no significant difference was observed in the 3rd and 4th sections in terms of the mixing ratio. This result indicates that the additive having a specific composition according to the present invention has a growth promoting effect.

[0063]

Example 5 A breeding test using rabbits was conducted. After introducing 15 male rabbits (variety: Japanese white breed) weighing 700 g to 800 g (five weeks old) and preliminarily rearing them for one week,
13 birds were set with 5 birds per plot and tested. Each animal was bred in a gauge, and feed and drinking water were freely taken and fed ad libitum. For the feed, a feed for rabbit rearing was used.
A mixture of the additives prepared by the above method was used. Three kinds of test feeds were prepared. Feed 1 was about 5% by weight of additive, feed 2 was about 10%, and feed 3 was about 30%.
, And an additive-free feed was prepared as feed 4 as a control.

The feeding period was three months (until the average body weight became about three times that at the start of feeding). As a result of feeding the additive of the present invention, there was no death in any of the administration groups,
In the test groups 2, 3 and 4 to which the additive according to the present invention was fed, the rabbit hair became glossier than the control group, and no abnormal change was observed in the symptom observation.

Table 2 shows the changes in body weight and the rate of increase in body weight.

[0066]

[Table 2]

As shown in Table 2, the increase was observed in the sections 2, 3 and 4 as compared to the control section, and no significant difference was observed in the sections 3 and 4 with respect to the mixing ratio. From these results, it was shown that the additive of the specific composition according to the present invention has a growth promoting effect.

[0068]

Example 6 First chicks (average weight: 45 g) of broiler breeds (arbor acres), male and female, 50 birds each, were preliminarily reared for 2 days, and divided into two sections so that the number of males and females was the same. (The average weight of each test group was 50 g.
Met). The test feed broiler standard feed for the first stage of fattening was fed for 4 weeks. The breeding environment was reared on butteries (incubated at 35 ± 2 ° C.) until 4 weeks of age. In the test plot, an additive (formulation 2) prepared by the method shown in Example 1 was mixed with the above-mentioned formula feed, and this was mixed with the broiler test feed so as to have a weight percentage of about 20%. Each of the broiler test feeds without additives was fed. The breeding was carried out with water supply freely by a water supply device, and the food was constantly fed and maintained in a state where the food remained constantly.

The results of the four-week feeding are shown in Table 3.

[0070]

[Table 3]

The weight gain index in Table 3 is calculated by assuming that each of the male and female in the control group is 100%. As is clear from the results in Table 10, as a result of feeding for 4 weeks, an increase in body weight of about 24% was observed as compared with the non-added control group. Sensory tests confirmed that the smell of broiler dung in the test plot was significantly lower than that in the control plot.

[0072]

EXAMPLE 7 Two-month-old piglets (landrace, 10 males and 10 females) each having an average body weight of 19 kg were divided into two groups, a test group and a control group. The method of raising pigs was as follows. The breeding environment was a windowless pig house in a private room, and ventilation was performed with a ventilation fan. The diet was raised on a standard diet for piglets.

The feed was constantly fed and the feed was constantly left in the feed box, and the water was supplied automatically by a water supply device. In the test plot, the additive (formulation 3) prepared in Example 3 was mixed at a weight ratio of about 20% to prepare a test feed, and a control pig-free feed was added as a control.

After breeding for 3 weeks under the above breeding conditions,
As shown in Table 4, the pigs in the test group fed with the additive according to the present invention showed a clear growth promoting effect as compared with the control group.
In the test group fed with the additive of the present invention, a significant decrease in fecal odor was also confirmed in the sensory test.

[0075]

[Table 4]

[0076]

Example 8 Approximately 500 yellowtail fry with an average fish weight of about 100 g
Two small fish net cages that breed 0 fish were used as a test plot and a control plot, respectively. The feeding period is 30 days, the control group is a moist pellet obtained by mixing a commercially available mixed feed for yellowtail and minced sardine at a ratio of 1: 1, and the test group is further supplemented with the additive according to the present invention (formulation 3). Feed 10
What was added so that it became about 10 g per 0 g was supplied. The water temperature during the test was 24-26 ° C.

Table 5 shows the results of the fish weight, food intake, and weight gain.

[0078]

[Table 5]

In the test group in which the additive of the present invention was added to the feed, the weight gain was superior to the control group compared to the control group.

3. Example of plant cultivation

[0081]

Example 9 Using the mixed culture solution prepared in Example 1,
Komatsuna cultivation test was conducted. The test conditions are as follows. A small plastic pot was filled with commercially available seedling soil, and the mixed culture was irrigated with the soil at a rate of 200 liters, 500 liters, and 1000 liters per 10 ares, and the control plot was irrigated with water. The seedlings were sown with Komatsuna, and a raising test was conducted for 33 days. The irrigation treatment was carried out on the 5th, 12th and 21st days after sowing under the same conditions as before sowing in each test section and control section. The test results are shown in Table 6.

[0082]

[Table 6]

[0083]

Example 10 As a culture solution, a mixed culture solution obtained by mixing a culture solution containing cells obtained by separately culturing cells coexisting in the culture solution was used. When carried out under the same conditions as in Examples 1 to 9, good results similar to those of the above examples were obtained.

[0084]

Example 11 Further, the same procedures as in Examples 9 to 9 were carried out using the co-culture medium filter obtained by filtering the cells from the co-culture medium used in each of the above-mentioned examples in a conventional manner. Under these conditions, good results similar to those of the above examples were obtained.

[0085]

Example 12 Further, using the mixed culture medium filter obtained by filtering the cells from the mixed culture medium used in Example 10 by an ordinary method, the above Examples 1 to 9 were used.
When carried out under the same conditions as in the above, good results similar to those of the above examples were obtained.

[0086]

According to the present invention, livestock can be safely and healthily promoted by feeding livestock with a co-culture liquid of lactic acid bacteria and yeast according to the present invention or an additive obtained by shaping the liquid. For example, pets such as dogs and cats, cows, pigs,
Livestock such as chickens, rabbits, deer, goats, monkeys, mice, rats,
It is widely effective for animals such as laboratory animals such as guinea pigs.

In particular, when used as a feed, it is possible to increase the taste by mixing about 10 to 20% with a normally fed feed to improve the appetite of cattle, pigs and other livestock animals themselves. The nutritive value of the feed is increased by the synergistic effect of the useful microorganisms added during the production, and the digestion function is enhanced by improving the digestion function, the digestibility of the roughage is also increased, and the digestion and absorption of nutrients is enhanced. Can be done.

In addition, the effective bacteria in the digestive system are multiplied to suppress harmful bacteria, and as a result, the fattening rate is improved, the meat quality is improved and improved, and the feed cost is reduced. There is. Furthermore, the odor of the livestock itself, which is the main cause of the generation of odors peculiar to the livestock production process, is reduced.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12R 1: 865) C12R 1: 865) (C12N 1/20 (C12N 1/20 E C12R 1: 245) C12R 1: 245) (C12N 1/20 (C12N 1/20 E C12R 1:23) C12R 1:23) (C12N 1/20 (C12N 1/20 C12R 1:25) C12R 1:25) (C12N 1/20 (C12N 1/20 C12R 1:46) C12R 1:46) (C12N 1/20 (C12N 1/20 C12R 1:01) C12R 1:01) (71) Applicant 599166529 El Grant & Sons, Inc. 68633, Nebraska, USA, 152 Dodge, Oak Street 152, P.O.Box 157 (72) Inventor Hidetaka Tokugawa 2077-2, 202F, Matsudo, Matsudo-shi, Chiba Over-time (reference) 2B022 AA01 AB01 AB11 AB13 AB15 2B150 AA01 AA02 AA03 AA05 AA08 AA10 AA20 AB01 AB02 AB04 AB05 AB12 AC01 AC02 AC03 AC05 AC06 AC07 AC11 AC24 AC25 AC26 DD11 DD26 4B065 AA30X AA49X AA80X BB12 BB14 BB22 BB40 BD07 CA43 CA49

Claims (9)

[Claims] 1. A bioactive agent comprising a culture solution obtained by co-culturing lactic acid bacteria and yeast, wherein said culture solution contains cells of lactic acid bacteria and yeast, respectively, and the unit of said culture solution is The ratio of the number of lactic acid bacteria to the number of yeast cells per amount is 100: 10
A bioactive agent characterized by being in the range of １００100: 30. 2. A bioactive agent comprising a mixed culture obtained by mixing cultures obtained by separately culturing lactic acid bacteria and yeasts, wherein the mixed cultures are cells of lactic acid bacteria and yeast cells, respectively. And the ratio of the number of lactic acid bacteria to the number of yeast cells per unit amount of the mixed culture solution is 100: 10 to 100: 3.
A bioactive agent characterized by being in the range of 0. 3. The culture medium according to claim 1 or the mixed culture medium according to claim 2, further comprising at least one or more bacterial cells selected from the group consisting of inorganic vegetative bacteria, photosynthetic bacteria, and bacteria of the genus Bacillus. A bioactive agent added, wherein the bioactive agent is
The cells are treated with 100 lactic acid bacteria per unit amount of the bioactive agent.
A bioactive agent, which is contained in the range of 10 to 50. 4. The vegetative bacterium according to claim 3, wherein the vegetative bacterium is Nitrobacter, Nitrosomonas (N).
itrosomonus, Thiobacillus (Thiob)
acillus) and Tiobacterium (Thio)
(Bacterium). A bioactive agent characterized by being a bacterium selected from the group of B. bacterium). 5. A method according to claim 1, wherein said culture solution or mixed culture solution further comprises Bacillus subtilis.
lis) and Bacillus mesentericus (Bacill)
The bioactive agent according to claim 1, wherein the cells are prepared by adding and mixing cells of L. mesentericus in the range of 20 to 40 and 10 to 20 with respect to 100 lactic acid bacteria, respectively. 6. The bioactive agent according to claim 1, wherein the lactic acid bacterium contains at least one bacillus lactic acid bacterium and at least one cocci lactic acid bacterium. 7. A bioactive agent comprising a culture filtrate obtained by removing cells from the culture solution or the mixed culture solution according to claim 1. 8. An animal feed additive obtained by shaping the bioactive agent according to any one of claims 1 to 7 with a shaping material. 9. An animal feed comprising the animal feed additive according to claim 8 in an amount of 10 to 30% by weight based on the feed.