WO2001080663A1

WO2001080663A1 - Animal feed additives and method of producing the same

Info

Publication number: WO2001080663A1
Application number: PCT/KR2001/000703
Authority: WO
Inventors: Ryo Kumazaki
Original assignee: Ryo Kumazaki
Priority date: 2000-04-27
Filing date: 2001-04-27
Publication date: 2001-11-01
Also published as: KR100541379B1; TW557201B; JP2001299230A; AU2001256802A1; KR20020093996A

Abstract

The present invention provides a method of producing animal feed additives which improve an environment of enteric bacteria, suppresses resistance bacteria and enhances the quality of animal products. That is, the present invention provides a technology for improving the environment of enteric bacteria of livestock to create a sound enteric environment, resulting in healthy excretions, compressing pathogenic bacteria, and enhancing animal product quality and productivity, thereby reducing expenses for managing a farm in general. The animal feed additives are prepared by adding an antioxidant and a viable cell agent to a high quality organic substance and fermenting the mixture in a sealed and air permeable container at a temperature range of from 25 °C to 37 °C for a certain period of time.

Description

ANIMAL FEED ADDITIVES AND METHOD OF PRODUCING THE SAME

Technical Field

The present invention relates to animal feed additives and a method of producing the same. In particular, the present invention relates to animal feed additives prepared by fermenting a mixture of a high quality organic substance, antioxidant and viable cell agent (preferably, a complex cultivation viable cell made of lactic acid bacteria and yeast, or a complex cultivation viable cell made of lactic acid bacteria, yeast and acetic bacteria, and photosynthetic bacteria) through antioxidation and fermentation of the viable cells. Background Art

Environmental problems related to the odor generated from livestock farms and poultry farms is a very serious problem for many neighbors, and farmers themselves. Therefore, some farmers had to live far away from other neighborhoods or tried an ozone generator hoping that it would somewhat reduce the odor. Sometimes, the farmers used different kinds of antibiotics in order to prevent any type of diseases of livestock. Unfortunately, this led to other problems, such as the creation of new resistant bacteria and putting the farmers in financial difficulties.

Disclosure of the Invention

It is, therefore, an object of the present invention to provide animal feed additives and a method for preparing the same, which improves an environment of enteric bacteria, suppresses resistance bacteria and enhances the quality of animal products. That is, the present invention provides a technology for improving the environment of enteric bacteria of livestock to create a sound enteric environment, making healthy excretions, suppressing pathogenic bacteria, and enhancing animal product quality and productivity, thereby reducing the expenses of managing a farm in general.

To achieve the above object, research has been conducted particularly on animal feed additives by fermenting the mixture of a high quality organic substance, antioxidant and viable cell agent (preferably, a complex cultivation viable cell made of lactic acid bacteria and yeast, or a complex cultivation viable cell made of lactic acid bacteria, yeast and acetic acid bacteria, and photosynthetic bacteria). The resulting animal feed additives contains plenty of amino acid, organic acid, vitamins and nucleic acid produced from proliferated viable cells. Thus, when the additive included in the animal feed is administered to animals, they form a very sound enteric condition due to the enteric bacteria grown inside of their bodies.

As mentioned before, the animal feed additives of the present invention is prepared by adding antioxidants and viable cell agents to a high quality organic substance and fermenting the mixture in a sealed and air permeable container at a temperature range of from 25°C to 37°C. Preferably, the antioxidant and the viable cell agent are used at an amount of 15 to 25% by weight of the organic substance.

According to a preparing method of an animal feed of the present invention, the antioxidant suppresses excessive oxidation of the organic substance and lactic acid, which is a product of lactic acid bacteria out of the viable cell agents, and controls saprophytic bacteria in the organic substance. In addition, oxygen is consumed by aerobic bacteria existing in air or being attached to material, aerobic bacteria (incipient bacteria) or facultative anaerobic bacteria contained in a viable cell agent. On the other hand, photosynthetic bacteria consume carbon dioxide produced by the above bacteria, and a great amount of carbon dioxide produced in turn suppresses excessive proliferation of aerobic bacteria or facultative anaerobic bacteria themselves. Therefore, although a thermal treatment is not carried out on a high quality organic substance, saprophytic bacteria in a sealed and permeable container, such as a plastic bag, can still be suppressed. In addition, plenty of nutrients for enteric bacteria such as, amino acid, organic acid, vitamins and nucleic acid are produced by proliferation of viable cell groups and the fermentation process. Best Mode for Carrying Out the Invention

According to a preferred embodiment of the present invention, antioxidant solution containing an antioxidant and a viable cell agent were added to a high quality organic substance and stirred together. Here, moisture content is adjusted to be in a range of from 15% to 25%) by weight. If the moisture content is not high enough, water should be added. But, if the moisture content is in the range, there is no need to add water.

Then, the mixture containing the high quality organic substance was put in a sealed and permeable container like a plastic bag at a temperature of 25°C to 37°C for a certain period of time and fermented to yield animal feed additives. The animal feed additives obtained is applied or spread to livestock. In general, the animal feed to feed animals contains approximately 0.4 wt% to 3wt%> of the animal feed additives.

The viable cell agent contains at least one kind of lactic acid bacteria, yeast and photosynthetic bacteria, respectively. Sometimes, acetic acid bacteria are optionally added. Particularly, lactic acid bacteria are used in combination with other kinds, preferably four different kinds of bacteria. In general, lactic acid bacteria are prepared by carrying out phagocytosis using freeze-dried mycobiont and culturing them in an appropriate medium to obtain culture (medium). In case of using plural kinds of lactic acid bacteria, each lactic acid bacterium should be cultured in a separate culture and later mixed with other culture prepared separately.

Similarly, yeast is prepared by carrying out phagocytosis using freeze-dried mycobiont and culturing them in an appropriate medium to obtain culture (medium). Although multiple kinds of yeast fungi can be applied together, it is more preferable to culture each kind of yeast in a separate culture and mix with other culture later. In addition, photosynthetic bacteria are prepared by carrying out phagocytosis using freeze-dried mycobiont (preferably more than 2 kinds) and culturing them in an appropriate medium to obtain culture (medium). In case of using plural kinds of photosynthetic bacteria, each photosynthetic bacterium should be cultured in a separate culture and later mixed with other culture prepared separately. Adding to the above, in case of using acetic acid bacteria additionally, the same method described above is applied here also. That is, acetic acid bacteria are prepared by carrying out phagocytosis using freeze-dried mycobiont and culturing them in an appropriate medium to obtain culture (medium). As described above, in order to prepare animal feed additives of the present invention, the mixture of a high quality organic substance, an antioxidant and a viable cell agent is fermented in a sealed container at 25°C to 37°C for a certain period of time.

As for the viable cell agent, culture prepared by lactic acid bacteria, yeast, acetic acid bacteria and photosynthetic bacteria are used, combined or separately.

A more preferable embodiment of the present invention introduces a slightly different method. In other words, in order to make animal feed additives, a complex culture is first prepared by mixing acetic acid bacteria culture and yeast culture or, optionally acetic acid bacteria culture, and then adding this culture to an antioxidant for the reaction in the presence of glucide. Later, the complex culture and the culture of photosynthetic bacteria were added to a high quality organic substance and the resultant was fermented to yield the animal feed additives.

The preparation method of the animal feed additives in accordance with the above- described embodiment, in which culture of lactic acid bacteria and that of yeast, or optionally that of acetic acid bacteria were mixed in advance and added to an antioxidant in the presence of glucide for a complex cultivation, is on the basis of the following details.

1 wt%> to 3 wt% of culture of lactic acid bacteria, each containing 10¹⁰ to 10ⁿ of lactic acid bacteria / iL, respectively, 1 wt% to 3 wt% of culture of yeast, each containing 10^s to 10⁹ of yeast fungi /mL, respectively, 1 wt% to 5 wt% of antioxidant matter containing 0.01 wt% to 0.05 wt% of antioxidant, and 3 wt%> to 5 wt% of glucide (preferably, molasses) are mixed all together to yield a complex culture that has designated pH (3.2 through 3.7) at a temperature range of from 25°C to 37°C. Optionally, 1 wt% to 2 wt% of culture of acetic acid bacteria containing 10¹⁰ to 10¹¹ of acetic acid bacteria /mL can be mixed with other culture. In addition, the present specification introduces the number of bacteria measured by using a plating method.

To a high quality of organic substance, was added the mixture of 5 wt% to 10 wt% of the complex culture, 5 wt% to 10 wt% of antioxidant solution containing 0.01 wt% to 0.05 wt% of antioxidant, and a culture containing 10⁸ to 10¹⁰ of photosynthetic bacteria /mL or a plurality of culture containing the same numbered photosynthetic bacteria, respectively.

Here, the mixing ratio of the high quality organic substance is adjusted so that moisture content of the mixture is within a range of from 15 wt% to 25 wt%>. The moisture content aforementioned can be designated by adding the organic substance and water additionally.

As explained before, the animal feed additives of the present invention are prepared by fermenting a mixture of a high quality substance and combination of a viable cell agent and an antioxidant in a sealed and permeable container, for example, a plastic bag, at a temperature range of from 25°C to 37°C for a certain period of time.

Then the animal feed additives prepared by the above method are applied or spread to livestock. Normally, the animals are provided with animal feed having 0.4 wt% to 3 wt% of animal feed additives.

The sealed and permeable container indicates a container that is shut off from outside in terms of air permeability, yet inside of the container, the air entered at the time of closing the container continuously circulates in the high quality organic substance. The time required for fermentation may vary depending on moisture content of the high quality organic substance after it is mixed with other addition, but usually one week to three weeks are sufficient. The high quality organic substance of the present invention is selected from a group consisting of starch containing substance, protein containing substance or fat containing substance, each being fresh and slow to oxidation and having moisture content below 13%.

Specifically, the organic substance includes at least one kind of a small amount of rice bran, fish dregs, cornstarch, sesame dregs, wheat bran or soybean dregs. In addition, as for an adsorbent of the odor generated by the high quality organic substance, activated carbon, fossil shellfish as a source of calcium, iron, manganese, copper, zinc, or a shell of a crab or conch as a source of molybdenum mineral.

The antioxidant is one of vitamin A, vitamin B group or vitamin E. Particularly, KMX of vitamin B group, manufactured by KORIN KOREA CO., is preferred. The glucide is selected from a group consisting of glucose, mannose, fructose, white sugar, maltose, cellobios, lactose, trehalose, melibiose, raffinose, esculin, salicyn, amygdalin, mannite, sorbit, sorbose or mentitose. Occasionally, molasses, saccharide or oligosaccharide is used in the industry.

On the other hand, as for the viable cell agent, more than one kind of basic photosynthetic bacteria, facultative basic lactose bacteria and yeast, respectively. Optionally, aerobic acetic bacteria can be added also. More preferably, the lactic acid bacteria are selected from a group consisting of lactobasillus paracasei subsp. paracasei, lactobacillus salivaius, lactobacillus acidophilus, lactobacillus deibrueckii subsp lactis, lactobacillus brevis or lactobacillus buchneri. The photosynthetic bacteria are selected from a group consisting of rhodopseudomonas pahstris and rhodopseuaomonas spheroids. The yeast is either saccaromyces cerevisiae or Candida valida. The acetic acid bacteria are selected from a group of acetobaccter liquefaciens. Accordingly, the animal feed additives of the present invention are very advantageous to livestock and a large number of farmers because the fermented animal feed additives promote proliferation of viable cells and the viable cells produce nutrients like amino acid, lactic acid, vitamins, and nucleic acid that help enteric bacteria of the animals to grow actively and creates a sound intestinal surrounding, so it is now possible to obtain healthy excretions of the animals, suppress pathogenic bacteria and odor, and enhance animal product quality and productivity.

It is to be understood that although particular embodiments are described in more detail for the purpose of explaining the effects of the animal feed additives of the present invention, the present invention is not intended to be limited to the embodiments only. Example 1

In Example 1, there is provided a method for preparing culture for different bacteria, respectively, required for preparing the animal feed additives of the present invention. Also, the effects of the animal feed additives are tested. Preparation of Culture for Lactic Acid Bacteria

As for the lactic acid bacteria to be used for a viable cell agent, four kinds of freeze- dried bacteria, named JCM1039, JCM1046, JCM1134 and JCM1248, were selected, which are described in the catalogue distributed by JCM Microorganism Week published by the microorganism preservation facilities of an institute of physical and chemical research. In order to make the culture (medium) having the pH from 6 to 6.5, lOg of peptone, lOg of meal extract, 5g of yeast extract, 20g of glucose, lg of twin 80, 2g of potassium dihydrogen phosphate, 5g of sodium acetate, 2g of citrate diammonium, 0.2g of magnesium sulfate hydrate, and 50mg of manganese sulfate hydrate were dissolved in 1 liter of distilled water. Then, the lactic acid bacteria selected from the catalogue were cultured in the culture according to the bulk starter method at 37°C and yielded 10¹⁰/mL of the culture. Preparation of Culture for Yeast Fungi

As for the yeast fungi to be used for a viable cell agent, two kinds of freeze-dried bacteria named JCM1499 and JCM3573 were selected, which were introduced in the catalogue distributed by JCM Microorganism Week published by the microorganism preservation facilities of an institute of physical and chemical research. The culture (medium) having the pH from 5.4 to 5.6 is prepared by dissolving 200g of potatoes, lOg of glucose, 15g of agar in 1 liter of distilled water. Then, the yeast fungi selected from the catalogue were cultured in the culture according to bulk starter method at 25°C and yielded 107mL of the culture.

Preparation of Culture for Acetic Acid Bacteria

As for the acetic acid bacteria to be used for a viable cell agent, freeze-dried bacteria named IF012388 was selected which were introduced in the catalogue distributed by IFO Microorganism Week published by a zymology research foundation. The culture (medium) having pH 6.8 is comprised of 5g of yeast extract, 3g of peptone, 30g of glucose, lOg of calcium carbonate, 15g of agar and 1 liter of distilled water. Then, the acetic acid bacteria selected from the catalogue were cultured in the culture at 30°C under the aerobic condition, and yielded 10⁹/mL of the culture. Preparation of Culture A for Photosynthetic Bacteria

As for the photosynthetic bacteria to be used for a viable cell agent, freeze-dried bacteria named JCM2524 were selected, which was introduced in the catalogue distributed by JCM Microorganism Week published by the microorganism preservation facilities of an institute of physical and chemical research. The culture (medium) having pH 7 is prepared by dissolving 2g of yeast extract, 2g of sodium L-malate, 2g of monosodium glutamate, lg of potassium hydrogen phosphate, 0.5g of sodium bicarbonate, 0.2g of magnesium sulfate heptahydrate, 0.2g of calcium chloride bihydrate, 2mg of manganese sulfate hydrate, 0.2g of ferrous sulfate heptahydrate, 0.5mg of cobalt chloride hexahydrate, Img of thymine-HCl, Img of nicotinicacid, O.lmg of biotin in 1 liter of distilled water. Then, the photosynthetic bacteria were cultured in the culture under a 2000Lux fluorescent lamp at 25°C according to the anaerobic method, and yielded 10⁹/mL of the culture. Preparation of Culture B for Photosynthetic Bacteria

As for another kind of photosynthetic bacteria to be used for a viable cell agent, freeze-dried bacteria named ATCC 17023 were selected, which was introduced in the catalogue called ATCCC Bacteria and Bacteriophages published by America Type Culture Collection. The culture (medium) having pH 6.9 is prepared by dissolving 2.5g of malic acid, lg of yeast extract, 2g of ammonium sulfate, 0.2g of magnesium sulfate heptahydrate, 0.07g of calcium chloride byhydrate, 0.0 lg of ferric citrate, 0.02g of ethylenediamine IV acetic acid, 0.6g of potassium hydrogen diphosphate, 0.9g of potassium dihydrogen phosphate, Img of microelement (prepared by dissolving 0.3g of ferric citrate, 0.002g of MnSO₄, 0.00 lg of H₃BO₃, 0.002g of (NH₄)₆Mo₇O₂₄ ^■ 4 H₂O, 0.05g of EDTA, 0.02g of CaCL, ^• 2H₂O i n 100ml of distilled water), 7.5ml of vitamin complex (prepared by dissolving 0.2g of nicotinicacid, 0.2g of nicotinamide, 0.4g of thymine-HCl, 0.008g of biotin in 1 liter of distilled water) in 1 liter of distilled water. Then, the photosynthetic bacteria were cultured in the culture under a 2000Lux fluorescent lamp at 30°C according to the anaerobic method, and yielded 10⁹/mL of the culture. Preparation of Complex Culture for Lactic Acid Bacteria, Yeast and Acetic Acid Bacteria

The complex culture is prepared by culturing 3 wt% of each culture for lactic acid bacteria, respectively, 3 wt% of each culture for yeast, respectively, 2 wt% of the culture for acetic acid bacteria, 10 wt% of KMX solution manufactured by KORTN KOREA CO., containing 0.01 wt%> of antioxidant, and 5 wt% of molasses as glucide at 27°C. As a result, the culture with pH 3.2 was obtained, which contained lactic acid and yeast as much as 10⁹ to 10¹⁰ per milliliter.

Then, 10 wt% of KMX solution manufactured by KORIN KOREA CO., LTD., containing 0.01 wt% of antioxidant, 10 wt% of the complex fluid comprised of each culture for the lactic acid bacteria, yeast and acetic acid bacteria, respectively, 5 wt% of each culture A and B, respectively, for the photosynthetic bacteria, 2 wt%> of fish dregs, 5 wt%> of wheat bran, 5 wt% of soybean dregs and 1 wt% of crab shell were mixed and stirred all together. After adjusting the moisture content to be approximately 22 wt%>, the resulting mixture was put in a plastic bag and fermented at 37°C for about 2 weeks to yield the animal feed additives of the present invention.

The fermented animal feed additives prepared by a method according to the present invention (hereinafter, it is abbreviated as "the fermented animal feed additives") was fed to the layers (i.e., eggs-laying hens) for a test. The breeding of the hens followed the general breeding management procedures, and the aviary for the hens was thoroughly disinfected in accordance with 'all-in all-out' method.

To conduct the test, first, a number of layers were purchased from an aviary. Then, to the animal feed particularly for the layers on the market, added was 3 wt% of the fermented animal feed additives. The animal feed was then distributed to a control zone for providing the hens with the animal feed additives through an automatic supplier, an object zone for providing the animal feed, and a cage for the layers. Each hen was given two tentative weeks for the test.

In addition, the amount of the remaining animal feed for each hen was measured every week in order to calculate the ingested amount of the animal feed. In this case, the ingestion percentage of the animal feed was also calculated on the basis of the total amount of the animal feed ingested by a hen. The percentage of laying eggs was calculated by comparing the total number of eggs collected at 10 o'clock every morning and the frequency of laying-egg by the hens in the control zone and the object zone. Then, 6 eggs were arbitrarily selected every day to measure total weight of each egg, weight of the yolk and weight of the eggshell by using an electronic scale. Using a caliper, the long diameter and short diameter of an egg and thickness of the eggshell were measured. Table 1 shows the results of productivity after the fermented additive was provided to the layers. <Table 1>

Based on the results shown in Table 1, the following facts can be induced:

1) The percentage of laying egg of hens in the control zone, 83 >, was increased by 3.75%, compared to that of the object zone, 80%.

2) The weight of an egg in the control zone, 66.05g, showed 3.67% of increase, compared to that of the object zone, 63.71g. If the increment 2.34g of each egg is multiplied by the total number of the eggs, the total increase is very significant.

3) The weight of a yolk of an egg in the control zone, 16.08g, showed 1.53% of decrease, comparing to that of the object zone, 16.33g. It is believed that decrease in cholesterol content in a yolk probably affected the total weight of the yolk.

4) The short diameter of an egg in the control zone showed 0.9%) of increase and the long diameter 5.93% of increase, compared to corresponding diameters of an egg in the object zone. It is believed that increased percentage of laying egg and the total weight of an egg probably affected the length of each diameter.

5) The thickness of an eggshell in the control zone showed 1.7%, 0.007mm to be more specific, of increase. Also, the weight of an eggshell showed 3.83% of increase, comparing to the eggs in the object zone. Thanks to the thickened eggshell in the control zone, fewer eggs were broken. The weight of the eggshell was increased because the eggshell became thicker and the egg itself was larger overall.

6) The percentage of broken eggs in the control zone, 1.4%, showed 22.2% of decrease, compared to that of the object zone, 1.8%. Fewer broken eggs were observed in the control zone because the eggshell got thicker and heavier.

Example 2

A study was conducted on changes of salmonella pathogenic bacteria. Similar to Example 1, the layers in the control zone and the object zone were divided into three groups and the changes of salmonella typhimuium inside the excretions of the hens were measured. The results are shown in Table 2. <Table 2>

(log _I0 salmonella gm excretion)

As shown in Table 2, the layers in the control zone had 59% fewer salmonella typhimuium, compared to the hens in the object zone. Example 3

Similar to Example 1, 15 of layers in the control zone and the object zone were divided into 3 groups according to their wings. Their fresh excretion was collected and cultivated. Then, the number of bacteria existing in the excretion and the number of occurrence frequency thereof were calculated in comparison with those of hens in the object zone. The results are shown in Table 3. <Table 3>

From to the test result of salmonella culture-positive chick/total%, it is known that occurrence frequency of the pathogenic bacteria by cultivation of the hens in the control zone is even less than a half of that of the hens in the object zone. Example 4

Similar to Example 1, the layers in the control zone and the object zone were divided into three groups and changes in ammonia content (μg/g) in their excretion were studied by using an ammonia analysis indicator. The results are shown in Table 4. <Table 4>

The results in Table 4 manifest that the layers in the control zone reduced the ammonia content down to 55% to 79%). When the intestinal substance of the hens in two zones was compared, the difference in the ammonia content was even greater. Example 5

Similar to Example 1, the layers in the control zone and the object zone were divided into three groups and changes in cholesterol content in their yolks were studied by using a measurement kit for cholesterol test. The results are shown in Table 5. Especially, the sample survey on the eggs of the layers fed with fermented animal feed additives lasted for 90 days. <Table 5>

As shown in Table 5, the cholesterol content in layers of the control zone was gradually decreased in contrast to that of the hens in the object zone. Example 6

Similar to Example 1, the poulards were divided into 2 zones: the control zone and the object zone. Then, a breeding experiment was conducted on the poulards in both zones. In other words, 3 wt% of the animal feed additives obtained in Example 1 was added to the animal feed on the market and supplied to the poulards in order to find out how the animal feed additives affect the poulards. The breeding method of the poulards complied with the guidelines for the general breeding.

For the experiment, the poulards (120 winds and poulards) were moved from a hatchery to a 1^st age group and stayed therein for the preliminary period. After the prelimnary period, in a 3^rd age group, the poulards in the control zone were fed with the animal feed containing the animal feed additives obtained in Example 1 for 5 weeks while the poulards in the object zone were fed with the animal feed on the market only. A growth rate of each poulard in both zones was closely observed at the same time (i.e., 10 am) every day. Particularly, the average values of gained weight (final weight - initial weight) of the poulards in both zones are shown in Table 6. <Table 6>

As shown in Table 6, the poulards in the control zone grew at a normal pace, gaining approximately 1.5kg at the end of 5 weeks. Example 7 Similar to Example 1, the poulards were divided into 2 zones: the control zone and the object zone for an experiment to find out how the animal feed additives affect enteric bacteria of the poulards. For the experiement, the poulards (120 winds and poulards) were moved from a hatchery to the 1^st age group and stayed there for the preliminary period. Later, in the 3^rd age group, the pouldards in the control zone were fed with the aniaml feed containing 3 wt% of the animal feed additives obtained in Example 1 for 5 weeks while the poulards in the object zone were fed with the animal feed on the market only.

After 4 weeks, some of the poulards were slaughtered and their bellies were cut very carefully in a way of not contaminating the inside. Their small intestines and appendixes were cut into 1cm, diluted with distilled water and dissolved in the agar medium. Later, the samples were cultured in a microorganixm medium for 24 hours at 36°C. Dividing colony forming unit (cfu) into 8 groups, the changes in enteric colon bacilli of the poulards in the control zone after 4 weeks were observed. The results are shown in Table 7. <Table 7>

As shown in Table 7, the poulards in the control zone had 50% fewer enteric colon bacilli in average. It is believed that the microorganism included in the animal feed additives created very useful enteric bacteria. Industrial Applicability

In conclusion, the animal feed additives of the present invention are very useful for improving the environment of enteric bacteria of livestock to create a sound enteric environment, resulting in healthy excretions, suppressing pathogenic bacteria, and enhancing animal product quality and productivity, thereby reducing managing expense of a farm in general.

Claims

1. A method of producing animal feed additives comprising the steps of: adding an antioxidant and a viable cell agent to a high quality organic substance; placing the mixture in a sealed and air permeable container; and fermenting the mixture at a temperature range of from 25°C to 37°C for a certain period of time to prepare the animal feed additives.

2. The method of claim 1, wherein the sealed and air permeable container is a plastic bag.

3. The method of claim 1 or claim 2, wherein the viable cell agent comprises at least one kind of lactic acid bacteria, yeast and photosynthetic bacteria, respectively.

4. The method of claim 3, wherein the lactic acid bacteria is selected from a group consisting of lactobasillus paracasei subsp. paracasei, lactobacillus salivaius, lactobacillus acidophilus, lactobacillus deibrueckii subsp lactis, lactobacillus brevis or lactobacillus buchneri; the photosynthetic bacteria being selected from a group consisting of rhodopseudomonas pahstris and rhodopseuaomonas spheroids; and the yeast being selected from a group of saccaromyces cerevisiae or Candida valida.

5. The method of claim 3 or 4, wherein the viable cell agent further comprises acetic acid bacteria.

6. The method of claim 5, wherein the acetic acid bacteria are selected from a group of acetobaccter liquefaciens.

7. The method according to one of claims 1 through 6, wherem a moisture content in the high quality organic substance containing the antioxidant and the viable cell agent is in a range of from 15 wt% to 25 wf%.

8. Animal feed additives prepared by the method according to any one of claims 1 through 7.

9. An animal feed for livestock, comprising 0.4 wt% to 3 wt% of the animal feed additives of claim 8.