CN102987058B - Method for producing efficient biological feed by using movable type fermentation and high-energy raw materials - Google Patents

Method for producing efficient biological feed by using movable type fermentation and high-energy raw materials Download PDF

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CN102987058B
CN102987058B CN201210495545.5A CN201210495545A CN102987058B CN 102987058 B CN102987058 B CN 102987058B CN 201210495545 A CN201210495545 A CN 201210495545A CN 102987058 B CN102987058 B CN 102987058B
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张有聪
郝永清
史彬林
任国军
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Abstract

The invention discloses a method for producing efficient biological feed by using movable type fermentation and high-energy raw materials. The method comprises the following steps of: smashing various fermentation substrate raw materials and then adding into a mixer, then inoculating mixed bacterium liquid and water to mix; and filling the fermentation substrate after finishing inoculating and mixing in a silica gel breathing membrane feed fermentation bag through an automatic metering and packing system to pack and seal, and fermenting for 72-120 hours at a constant temperature in a fermentation chamber to obtain the finished product of the efficient biological feed. The fermentation substrate consists of the following components in parts by weight: 5-10 parts of oil bran, 30-40 parts of high-fat palm meal, 40-50 parts of primverose protein, 5-10 parts of high-fat flax meal and 10-20 parts of high-fat alcohol meal. The mixed bacterium liquid is prepared by mixing bacterium liquids of trichosporon capitatum, yarrowia lipolytica and geobacillus stearothermophilus.

Description

A kind of method that adopts portable fermentation and ENERGETIC MATERIALS to produce highly effective biological feed
Technical field
The invention relates to a kind of method that adopts portable fermentation and ENERGETIC MATERIALS to produce highly effective biological feed, belong to additive for microbe feedstuff field.
Background technology
The ratio maximum (being greater than 50%) of the common shared diet of energy feed, so it occupies very important status in diet.Energy feed can be divided into cereals, the real processed side product class of paddy, piece root, stem tuber and processed side product, Dairy Processing byproduct and all kinds of animal and plant fats etc.Generally all there is the shortcomings such as essential fatty acid and fat-soluble avitaminosis, crude fat content is lower, aliphatic acid forms imbalance, cost is higher, digestive utilization ratio is poor in these conventional energy feeds.Livestock and poultry are because production performance improves constantly, more and more high to the requirement of daily ration nutrient density especially Diet energy density, with conventional feed, are difficult to make high-energy diet.In view of these reasons, the energy feed product that utilized in recent years solid microbe fermentation technology to produce because its essential fatty acid content is higher, protein energy ratio balanced, digestive utilization ratio high, is widely used in livestock and poultry cultivation.Solid microbe fermentation technology is also in constantly innovation and development simultaneously, utilize the example of microorganism fodder fermentation to produce biological feed a lot, but the overwhelming majority is thick-layer solid-state fermentation technology, not only equipment investment is large, complex process, and very easily influenced by ambient temperature, constant product quality is poor.The birth of efficient respiratory membrane solid-state fermentation technology, has fundamentally solved this problem.Efficient respiratory membrane solid-state fermentation technology is a kind of life breath device that can make microorganism beneficial bacterium keep high activity growth metabolism under nature, it can make viable bacteria can keep for a long time high activity under field conditions (factors), has solved dexterously microorganism solid fermentation heat radiation, anaerobism control, packing, accumulating and the stability difficult problem in whole process of producing product.Efficient respiratory membrane solid state fermentation production technology is to using various light industry accessory substances or the assorted dregs of rice as fermentation substrate, raw material does not need through sterilization, adopt the efficient respiratory membrane of microorganism of original creation and first pack the portable microorganism solid fermentation technique of after fermentation, the compound culture of beneficial bacterium microorganism of production high energy, high protein.This technology production technology is simple, and cost is low, and small investment is workable, and finished product is easy to use, is particularly suitable for the use of field, vast rural culture (family).This invention is striven grain and feed safety problem to solving people and animals, builds a conservation-minded society and is significant.
Summary of the invention
The object of the invention is to, according to above-mentioned situation, provide a kind of method that adopts portable fermentation and ENERGETIC MATERIALS to produce highly effective biological feed.
For achieving the above object, the present invention is by the following technical solutions:
A method that adopts portable fermentation and ENERGETIC MATERIALS to produce highly effective biological feed, is characterized in that,
(1) will be various fermentation substrate raw materials add in mixer after pulverizing, then access mixed bacteria liquid and water mixes;
(2) the complete fermentation substrate of inoculation mixing is loaded in silica gel respiratory membrane feed fermentation bag and is packed, seals by automatic gauge packaging system, and ferment at constant temperature 72～120h in fermenting cellar, obtains described highly effective biological feed finished product;
Consisting of of described fermentation substrate: oily chaff 5～10 weight portions, high fat palm kernel meal 30～40 weight portions, primverose albumen 40～50 weight portions, high fat flax dregs of rice 5～10 weight portions, the high fat alcohol dregs of rice 10～20 weight portions;
Described mixed bacteria liquid is mixed by the bacterium liquid of head trichosporon cutaneum (Trichosporon capitatum), the sub-sieve yeast (Yarrowia lipolytica) of solution fat and stearothermophilus ground bacillus (Geobacillusstearothermophilus).
Method as above, preferably, in step (1), described fermentation substrate is crossed 40～42 mesh sieves after pulverizing, mixed bacteria liquid is pressed the percentage by weight inoculation of fermentation substrate 5～10%, and water adds in the ratio of material-water ratio 1:1～1.5, and fermentation substrate moisture is adjusted between 45～55%.
Method as above, preferably, in step (2), in described packing, the process of sealing, that air in bag is not emptying, packaged material is 35～37 ℃ of constant temperature bottom fermentation 72～120h in fermenting cellar.
Method as above, preferably, described head trichosporon cutaneum is conciliate the sub-sieve yeast liquid of fat and is made by the following method:
Prepare first order seed culture medium: 12Brix brewer's wort 1000mL, sterilizing 20～30min at 115～121 ℃ of temperature;
Wherein the preparation method of brewer's wort is: fructus hordei germinatus is some, after pulverizing, adds 4 times to the water of malt weight, stirs, saccharification 4h in 55～60 ℃ of incubators, takes out and filters, and obtains brewer's wort, measure after the volume and concentration of this brewer's wort, be sub-packed in sterilized triangular flask 0.6kg/cm 2sterilizing 40min is standby, and the used time is diluted to required pol by filtrate;
Preparation secondary seed medium and fermentation medium: (NH 4) 2sO 42.0kg, glucose 10kg, yeast soak powder 5kg, MgSO 47H 2o0.5kg, KH 2pO 41.0kg, 10 ° of BX molasses 1000L, pH nature, sterilizing 20～30min at 115～121 ℃ of temperature;
Wherein the processing method of molasses is: the molasses stoste of 85 ° of BX uses water as 1:1.5 dilution.H 2sO 4adjust pH3.5, shake up, standing 8h, gets filtrate and adds milk of lime, adjusts pH7.2, and 60 ℃ are heated and stir 30min, and standing 12h, gets filtrate, adjusts pH6.0, is diluted to 10 ° of BX standby;
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane inoculated in first order seed culture medium in the ratio of ring/50～100mL, at 25～28 ℃ with 150～160r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 5～10% weight ratio, under the condition that is 1:1～1.5, with 140～150r/min mechanical agitation, cultivates 24～48h at 25～28 ℃, ventilation volume ratio;
Liquid fermentation and culture: the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 5～10% weight ratio, under the condition that is 1:1～1.5 at 25～28 ℃, ventilation volume ratio, with 140～150r/min mechanical agitation, cultivates 24～48h.
Method as above, preferably, described stearothermophilus ground bacillus bacterium liquid is made by the following method:
Prepare first order seed culture medium: peptone 5g, beef extract 3g, NaCl5g, MnSO 4h 2o0.005g, distilled water 1000mL mixes, sterilizing 20min～30min at 115～121 ℃ of temperature;
Preparation secondary seed medium and fermentation medium: soy meal 10kg, peptone 5kg, glucose 10kg, beef extract powder 2kg, MnSO 4h 2o5g, sterile pure water 1000L, pH nature, sterilizing 20～30min at 115～121 ℃ of temperature;
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane inoculated in first order seed culture medium in the ratio of ring/50～100mL, at 37～40 ℃ with 120～130r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 5～10% weight ratio, under the condition that is 1:0.8～1.2 at 37～40 ℃, ventilation volume ratio, with 120～130r/min mechanical agitation, cultivate 24～48h;
Liquid fermentation and culture: the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 5～10% weight ratio, under the condition that is 1:0.8～1.2 at 37～40 ℃, ventilation volume ratio, with 120～130r/min mechanical agitation, cultivates 24～48h;
Method as above, preferably, in the mixed bacteria liquid described in step (1), the mixed proportion of each bacterial classification and the inoculum concentration of mixed bacteria liquid are respectively:
The mixed proportion of head trichosporon cutaneum, the sub-sieve yeast of solution fat, stearothermophilus ground bacillus is 3～4:3～4:2～4, and inoculum concentration is 5～10% of fermentation substrate percentage by weight.
The highly effective biological feed that method prepares as mentioned above.
The highly effective biological feed that method prepares as mentioned above, wherein crude fat (DM)>=22.00%, crude protein (DM)>=20.12%, lipase activity>=13.70U/g, leukotrienes>=1.20%, linoleic acid>=8.50%, oleic acid>=9.30%, head trichosporon cutaneum (CFU/g)>=3.50 * 10 8, separate the sub-sieve yeast (CFU/g)>=5.00 * 10 of fat 8, stearothermophilus ground bacillus (CFU/g)>=8.0 * 10 8.
Beneficial effect of the present invention is:
The present invention is that to select oily chaff, high fat palm kernel meal, primverose albumen, high fat flax dregs of rice and the high fat alcohol dregs of rice be raw material, through pulverizing, add after water mixing, inoculate after the mixed bacteria liquid being formed by head trichosporon cutaneum (Trichosporoncapitatum), the sub-sieve yeast (Yarrowialipolytica) of solution fat, stearothermophilus ground bacillus (Geobacillus stearothermophilus) product of microorganism fermented forage making by efficient respiratory membrane process for solid state fermentation.The biological feedstuff crude fat content that adopts the inventive method production to obtain is high, and essential fatty acid content is abundanter.Simultaneously, the present invention adopts efficient respiratory membrane solid-state fermentation technology, can make microorganism beneficial bacterium under nature, keep high activity growth metabolism, solve dexterously microorganism solid fermentation heat radiation, anaerobism control, packing, accumulating and the stability difficult problem in whole process of producing product.
More particularly, the method for the portable fermentation process of employing provided by the invention and ENERGETIC MATERIALS production highly effective biological feed has the following advantages:
1, the inventive method adopts the solid-state fermentation technique of movable type of first packing after fermentation, and technological design has novelty, easy and simple to handle.
2, the efficient respiratory membrane bascule that the inventive method is used has creatively solved the technical barrier such as gas control and living contaminants in fermentation production process, easy and simple to handle controlled.During the fermentation, microbes produces the gases such as carbon dioxide, makes the air pressure in bag be greater than extraneous normal pressure.When bag internal gas pressure reaches a certain critical value, gas is discharged to the external world by this efficient respiratory membrane.The fermentation system but extraneous gas has no chance to enter all the time, disturbs thereby also just got rid of extraneous miscellaneous bacteria.
3, the specific microorganism fungus kind combination of the inventive method research and development can make beneficial bacterium occupy predominance, the ingenious advantage that combines anaerobic bacteria and aerobic bacteria fermentation, and raw material does not need sterilization just can be directly used in inoculated and cultured.
4, the inventive method can extensively utilize the agricultural of the high moisture contents such as bean dregs, pomace, corn steep liquor and sugared slag and light industry accessory substance as raw materials for production, production technology is practical, and added value of product is high, long shelf-life, tally with the national condition, be particularly suitable for promoting the use of in the vast rural area of China.
5, adopt the inventive method, the higher vegetalitas energy feed of crude fat content making, after the lipase degraded producing through microorganism solid fermentation, effectively improved the content of crude fat and the content of essential fatty acid in feed, the use value of feed is significantly improved.
6, adopt in the highly effective biological feed of the inventive method production crude fat (DM)>=22.00%, crude protein (DM)>=20.12%, lipase activity>=13.70U/g, leukotrienes>=1.20%, linoleic acid>=8.50%, oleic acid>=9.30%, head trichosporon cutaneum (CFU/g)>=3.50 * 10 8, separate the sub-sieve yeast (CFU/g)>=5.00 * 10 of fat 8, stearothermophilus ground bacillus (CFU/g)>=8.0 * 10 8.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of the inventive method.
The specific embodiment
By specific embodiment, describe technology of the present invention and feature in detail below, but these embodiment are not in order to limit protection scope of the present invention.
In following examples of the present invention, head trichosporon cutaneum used (Trichosporon capitatum), the sub-sieve yeast (Yarrowia lipolytica) of solution fat and stearothermophilus ground bacillus (Geobacillusstearothermophilus) bacterial classification are the wild-type strains of buying respectively in Chinese industrial microorganism fungus kind preservation administrative center (CICC) and Chinese common micro-organisms culture presevation administrative center (CGMCC).Preserving number is respectively: CGMCC2.1385, CICC1444, CICC21077.
Embodiment 1
Referring to Fig. 1, prepare in accordance with the following methods the highly effective biological feed of the present embodiment:
1. accurately take oily chaff 50kg, high fat palm kernel meal 300kg, primverose albumen 500kg, high fat flax dregs of rice 50kg, high fat alcohol dregs of rice 100kg, pour in mixer, then mixed bacteria liquid is inoculated by the percentage by weight of fermentation substrate 10%, water adds in the ratio of material-water ratio 1:1.2, mix continuously 3min, fermentation substrate moisture is adjusted to 50% left and right.
2. the fermentation substrate that inoculation mixes is loaded in efficient respiratory membrane feed fermentation bag and is packed, seals by automatic gauge packaging system, in packaging process, do not need air in bag emptyingly, packaged material 35 ℃ of condition bottom fermentation 96h in fermenting cellar are finished product.
3. the mixed bacteria liquid described in is to mix after head trichosporon cutaneum (Trichosporon capitatum), the sub-sieve yeast (Yarrowia lipolytica) of solution fat and stearothermophilus ground bacillus (Geobacillusstearothermophilus) are increased respectively to bacterium cultivation and liquid state fermentation by the following method, specifically:
(1) head trichosporon cutaneum (Trichosporon capitatum) is conciliate the sub-sieve yeast (Yarrowialipolytica) of fat:
A. prepare first order seed culture medium: 12Brix brewer's wort 1000mL, sterilizing 20min at 115 ℃ of temperature.
The preparation of brewer's wort: fructus hordei germinatus is some, after pulverizing, adds 4 times to the water of malt weight, stirs, saccharification 4h in 55～60 ℃ of incubators, takes out and filters, and obtains brewer's wort, measure after the volume and concentration of this brewer's wort, be sub-packed in sterilized triangular flask 0.6kg/cm 2sterilizing 40min is standby, and the used time is diluted to required pol by filtrate.
B. prepare secondary seed medium and fermentation medium: (NH 4) 2sO 42.0kg, glucose 10kg, yeast soak powder 5kg, MgSO 47H 2o0.5kg, KH 2pO 41.0kg, 10 ° of BX molasses 1000L, pH nature, sterilizing 30min at 115 ℃ of temperature.
The processing of molasses: molasses stoste (85 ° of BX) uses water as 1:1.5 dilution.H 2sO 4adjust pH3.5, shake up, standing 8h, gets filtrate and adds milk of lime, adjusts pH7.2, and 60 ℃ are heated and stir 30min, and standing 12h, gets filtrate, adjusts pH6.0.Be diluted to 10 ° of BX standby.
C. condition of culture:
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane is inoculated in first order seed culture medium in the ratio of ring/100mL, 28 ℃, 150r/min, shaken cultivation 24h.
Secondary seed is cultivated: the 10L automated seed canned secondary seed medium 5L that ferments, the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 10% weight ratio, and 28 ℃, ventilation volume ratio are 1:1.2, mechanical agitation 150r/min, cultivate 24h.
Liquid fermentation and culture: the 100L automated seed canned fermentation medium 50L that ferments, the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 10% weight ratio, 28 ℃, ventilation volume ratio are 1:1.2, mechanical agitation 150r/min, cultivate 24h.
(2) stearothermophilus ground bacillus
A. prepare first order seed culture medium: peptone 5g, beef extract 3g, NaCl5g, MnSO 4h 2o0.005g, distilled water 1000mL mixes, sterilizing 20～30min at 115～121 ℃ of temperature.
B. prepare secondary seed medium and fermentation medium: soy meal 10kg, peptone 5kg, glucose 10kg, beef extract powder 2kg, MnSO 4h 2o5g, sterile pure water 1000L, pH nature, sterilizing 20～30min at 115～121 ℃ of temperature.
C. condition of culture:
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane inoculated in first order seed culture medium in the ratio of ring/50mL, at 37 ℃ with 130r/min shaken cultivation 24h.
Secondary seed is cultivated: the 10L automated seed canned secondary seed medium 5L that ferments, the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 10% weight ratio, and 37 ℃, ventilation volume ratio are 1:1, mechanical agitation 130r/min, cultivate 24.
Liquid fermentation and culture: the 100L automated seed canned fermentation medium 50L that ferments, the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 10% weight ratio, 37 ℃, ventilation volume ratio are 1:1, mechanical agitation 130r/min, cultivate 24h.
4. in the mixed bacteria liquid described in, the inoculation mixed proportion of bacterial classification and the inoculum concentration of mixed bacteria liquid are respectively:
The mixed proportion of head trichosporon cutaneum, the sub-sieve yeast of solution fat, stearothermophilus ground bacillus is 4:4:2, and the inoculum concentration of mixed bacteria liquid is 10% of fermentation substrate weight.
5. the highly effective biological feed preparing according to above method, records crude fat (DM)>=24.73%, crude protein (DM)>=22.56%, lipase activity>=15.00U/g, leukotrienes>=1.20%, linoleic acid>=10.00%, oleic acid>=10.00%, head trichosporon cutaneum (CFU/g)>=5.00 * 10 8, separate the sub-sieve yeast (CFU/g)>=6.00 * 10 of fat 8, stearothermophilus ground bacillus (CFU/g)>=8.00 * 10 8.
Embodiment 2
Referring to Fig. 1, prepare in accordance with the following methods the highly effective biological feed of the present embodiment:
1. accurately take oily chaff 100kg, high fat palm kernel meal 300kg, primverose albumen 400kg, high fat flax dregs of rice 100kg, high fat alcohol dregs of rice 100kg, pour in mixer, then mixed bacteria liquid is inoculated by the percentage by weight of fermentation substrate 10%, water adds in the ratio of material-water ratio 1:1.2, mix continuously 3min, fermentation substrate moisture is adjusted to 50% left and right.
2. the fermentation substrate that inoculation mixes is loaded in efficient respiratory membrane feed fermentation bag and is packed, seals by automatic gauge packaging system, in packaging process, do not need air in bag emptyingly, packaged material 35 ℃ of condition bottom fermentation 96h in fermenting cellar are finished product.
3. the mixed bacteria liquid described in is by head trichosporon cutaneum (Trichosporon capitatum), separates the sub-sieve yeast (Yarrowia lipolytica) of fat and stearothermophilus ground bacillus (Geobacillusstearothermophilus) and by method as described in Example 1, after increasing bacterium cultivation and liquid state fermentation, mix respectively.
4. in the mixed bacteria liquid described in, the inoculation mixed proportion of bacterial classification and the inoculum concentration of mixed bacteria liquid are respectively:
The mixed proportion of head trichosporon cutaneum, the sub-sieve yeast of solution fat, stearothermophilus ground bacillus is 3:3:4, and the inoculum concentration of mixed bacteria liquid is 10% of fermentation substrate weight.
5. the highly effective biological feed preparing according to above method, records crude fat (DM)>=22.00%, crude protein (DM)>=20.12%, lipase activity>=13.70U/g, leukotrienes>=1.60%, linoleic acid>=8.50%, oleic acid>=9.30%, head trichosporon cutaneum (CFU/g)>=3.50 * 10 8, separate the sub-sieve yeast (CFU/g)>=5.00 * 10 of fat 8, stearothermophilus ground bacillus (CFU/g)>=9.50 * 10 8.

Claims (6)

1. a method that adopts portable fermentation and ENERGETIC MATERIALS to produce highly effective biological feed, is characterized in that,
(1) will be various fermentation substrate raw materials add in mixer after pulverizing, then access mixed bacteria liquid and water mixes;
(2) the complete fermentation substrate of inoculation mixing is loaded in silica gel respiratory membrane feed fermentation bag and is packed, seals by automatic gauge packaging system, and ferment at constant temperature 72～120h in fermenting cellar, obtains described highly effective biological feed finished product;
Consisting of of described fermentation substrate: oily chaff 5～10 weight portions, high fat palm kernel meal 30～40 weight portions, primverose albumen 40～50 weight portions, high fat flax dregs of rice 5～10 weight portions, the high fat alcohol dregs of rice 10～20 weight portions;
Described mixed bacteria liquid is mixed by the bacterium liquid of head trichosporon cutaneum, the sub-sieve yeast of solution fat and stearothermophilus ground bacillus;
Wherein, in mixed bacteria liquid described in step (1), the mixed proportion of each bacterial classification and the inoculum concentration of mixed bacteria liquid are respectively: the mixed proportion of head trichosporon cutaneum, the sub-sieve yeast of solution fat, stearothermophilus ground bacillus is 3～4:3～4:2～4, and inoculum concentration is 5～10% of fermentation substrate percentage by weight.
2. method according to claim 1, it is characterized in that, in step (1), described fermentation substrate is crossed 40～42 mesh sieves after pulverizing, mixed bacteria liquid is pressed the percentage by weight inoculation of fermentation substrate 5～10%, water adds in the ratio of material-water ratio 1:1～1.5, and fermentation substrate moisture is adjusted between 45～55%.
3. method according to claim 1, is characterized in that, in step (2), in described packing, the process of sealing, that air in bag is not emptying, packaged material is 35～37 ℃ of constant temperature bottom fermentation 72～120h in fermenting cellar.
4. method according to claim 1, is characterized in that, described head trichosporon cutaneum is conciliate the sub-sieve yeast liquid of fat and made by the following method:
Prepare first order seed culture medium: 12Brix brewer's wort 1000mL, sterilizing 20～30min at 115～121 ℃ of temperature;
Wherein the preparation method of brewer's wort is: fructus hordei germinatus is some, after pulverizing, adds 4 times to the water of malt weight, stirs, saccharification 4h in 55～60 ℃ of incubators, takes out and filters, and obtains brewer's wort, measure after the volume and concentration of this brewer's wort, be sub-packed in sterilized triangular flask 0.6kg/cm 2sterilizing 40min is standby, and the used time is diluted to required pol by filtrate;
Preparation secondary seed medium and fermentation medium: (NH 4) 2sO 42.0kg, glucose 10kg, yeast soak powder 5kg, MgSO 47H 2o0.5kg, KH 2pO 41.0kg, 10 ° of BX molasses 1000L, pH nature, sterilizing 20～30min at 115～121 ℃ of temperature;
Wherein the processing method of molasses is: the molasses stoste of 85 ° of BX uses water as 1:1.5 dilution, H 2sO 4adjust pH3.5, shake up, standing 8h, gets filtrate and adds milk of lime, adjusts pH7.2, and 60 ℃ are heated and stir 30min, and standing 12h, gets filtrate, adjusts pH6.0, is diluted to 10 ° of BX standby;
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane inoculated in first order seed culture medium in the ratio of ring/50～100mL, at 25～28 ℃ with 150～160r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 5～10% weight ratio, under the condition that is 1:1～1.5, with 140～150r/min mechanical agitation, cultivates 24～48h at 25～28 ℃, ventilation volume ratio;
Liquid fermentation and culture: the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 5～10% weight ratio, under the condition that is 1:1～1.5 at 25～28 ℃, ventilation volume ratio, with 140～150r/min mechanical agitation, cultivates 24～48h.
5. method according to claim 1, is characterized in that, described stearothermophilus ground bacillus bacterium liquid is made by the following method:
Prepare first order seed culture medium: peptone 5g, beef extract 3g, NaCl5g, MnSO 4h 2o0.005g, distilled water 1000mL mixes, sterilizing 20min～30min at 115～121 ℃ of temperature;
Preparation secondary seed medium and fermentation medium: soy meal 10kg, peptone 5kg, glucose 10kg, beef extract powder 2kg, MnSO 4h 2o5g, sterile pure water 1000L, pH nature, sterilizing 20～30min at 115～121 ℃ of temperature;
First order seed is cultivated: in 1000mL triangular flask, fill first order seed culture medium 500mL, strain inclined plane inoculated in first order seed culture medium in the ratio of ring/50～100mL, at 37～40 ℃ with 120～130r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid of cultivating through described first order seed is inoculated in secondary seed tank according to 5～10% weight ratio, under the condition that is 1:0.8～1.2 at 37～40 ℃, ventilation volume ratio, with 120～130r/min mechanical agitation, cultivate 24～48h;
Liquid fermentation and culture: the bacterium liquid of cultivating through described secondary seed is inoculated in fermentation tank according to 5～10% weight ratio, under the condition that is 1:0.8～1.2 at 37～40 ℃, ventilation volume ratio, with 120～130r/min mechanical agitation, cultivates 24～48h.
6. the highly effective biological feed preparing according to method described in any one in claim 1～5.
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