CN103109981A - Method for preparing feed additive by using bacillus licheniformis - Google Patents
Method for preparing feed additive by using bacillus licheniformis Download PDFInfo
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- 238000000855 fermentation Methods 0.000 claims abstract description 63
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Abstract
The invention provides a method for preparing a feed additive by using bacillus licheniformis. The method comprises the following steps of: preparing and activating a strain, preparing a strain with an inclined surface, preparing a triangular flask liquid strain, fermenting a canned solid and the like so as to obtain the feed additive. By adopting a unique culture medium and a solid fermentation technique, the strain propagation speed is accelerated, the fermentation period is shortened, the production cost is lowered, and the spore forming rate of a product is increased; a cheap medium such as bran is used as a main culture substrate, so that the production cost is effectively lowered; the production process has the unique advantages of water conservation, energy conservation, small quantity of byproducts and environment friendliness; and the production cost is low, the product is resistant to high temperature, acid, alkali and extrusion, is small in damage in the preparation process of the feed, small in adding quantity and remarkable in feeding effect, so that the method belongs to a sanitary production technique and has large-scale popularization and application values.
Description
Technical field
The present invention relates to feeding feed additive field, be specifically related to utilize bacillus licheniformis to prepare the method for feed addictive.
Background technology
The raiser cultivation during domestic birds and animals in order to reach prophylactic purpose, often add a large amount of antibiotic in feed, consequently inevitably cause abuse of antibiotics, produce in animal body medicament residue, finally accumulate the serious threat human health in human body by food chain.Studies show that bacillus licheniformis is all class probios, add in feed and can function as follows with its preparation of making: 1. bacillus is a kind of aerobic beneficial bacterium, it can form spore in adverse environment, high temperature resistant, acid and alkali-resistance and variations in temperature can be brought back to life rapidly after entering enteron aisle; And can consume rapidly limited oxygen in environment by fighting for limited nutriment with pathogenic bacteria, be conducive to keep the intestinal flora balance and play an important role, thereby suppress the pathogenic entero becteria growth, reach the effect of prophylactic treatment livestock and poultry intestinal canal diseases; 2. bacillus can produce multiple digestive ferment, and the help animal digests and assimilates nutriment, thereby promotes digestion and improve efficiency of feed utilization, reduces feeding cost; 3. bacillus can promote the animal intestinal associated lymphoid tissue to be in " the immune SBR " of height, and Development of Immune Organs is accelerated, and immune system is ripe fast and early, T, bone-marrow-derived lymphocyte quantity increase, and animal body fluid and cellular immune level improve; 4. bacillus growth and breeding in animal intestinal, can produce multiple nutrients material such as vitamin, amino acid, organic acid, somatomedin etc., participates in animal body metabolism, for body provides nutriment.
because bacillus preparation has above-mentioned important excellent characteristic, therefore attracted numerous Chinese scholars that it is furtherd investigate, research contents mainly concentrates on: the screening (Yang Xiaobin of bacillus, Xie Yongkui, Wei Yanpeng etc., the screening of Bacillus Subtilis Natto of Probiotics, Guangzhou Food Industry science and technology, supplementary issue in 2003), the preparation fibrinolysin (protect by complementary work, Guo Ailing, Qin Qiaoling etc., one plant height produces the screening of Nattokinase bacterial strain and the optimization of fermentation condition thereof, Food Science, the 4th phase in 2007), deep liquid fermentation process (Zhu Jianhui, Du Lianxiang, Lu Fuping, the Nattokinase liquid-state fermentation technology is optimized, food and fermentation industries, the 8th phase in 2005), pulvis preparation technology (Zhong Qingping, Wang Faxiang, Liu Jiachang, the development of Bacillus natto viable bacteria drying agent, food industry science and technology, the 8th phase in 2005) aspect such as, and concrete solid fermentation process has no report.
The tolerances such as the advantage that solid fermentation method is compared the liquid fermentation maximum is exactly that production cost is low, raw material simple, the product inulinase-producing activity is high, high temperature resistant, the acid and alkali-resistance of product, anti-extruding are strong.But the solid-fermented technique difficulty is large, is not easy to produce high-quality bacillus preparation.The preparation viable bacteria content that the production technology of present bacillus preparation and fermentative medium formula are produced is not high, and the preparation term of validity is short, makes product be difficult to prevention from suffering from the diseases and the growth-promoting effect that reaches desirable.
Summary of the invention
Content of the present invention is, for the prior art deficiency, provides a kind of solid fermentation cultural method that utilizes bacillus licheniformis to prepare feed addictive, the feed addictive viable bacteria content that the method makes is high, and the miscellaneous bacteria rate is low, and the brood cell is active strong, stability is strong, and production cost is not high.
The technical solution used in the present invention is that a kind of method of utilizing bacillus licheniformis to produce feed addictive is characterized in that comprising the steps:
(1) bacterial classification preparation and activation
The bacterial classification of the bacillus licheniformis of buying is made suspension, and adding sterilized water to be diluted to concentration is 10
-8The time, on the solid plate culture medium, it being coated with and separating or the line separation, the single bacterium colony of picking part carries out the mensuration of production capacity; Through repeated screening, determine the active bacterial strain strong, that viable count is high of the gemma that produces through fermentation and replace original bacterial strain.
(2) slant strains is made
By formula requirement configuration slant medium, the each component weight portion of slant medium is: beef extract 5 g, and peptone 10 g, NaCl 5 g, agar powder 20-25 g adds water and is settled to 1000 mL; PH is controlled at 7.2-7.4;
Each component is fully dissolved the slant medium heating, mix up pH, then be sub-packed in some test tubes, the sealing test tube; Test tube in 121 ℃ of sterilization 30 min, is taken out and puts the inclined-plane, cooling rear standby;
Above-mentioned slant medium is placed in drying box or incubator places the bacterial classification that 24-48h access step (1) prepares, wherein the bacillus licheniformis inoculum concentration be in test tube the mixture total weight amount 1%, cultivate 24h under 37 ℃ of conditions, the bevel bacterial classification.
(3) the triangular flask liquid spawn is made
By formula requirement configuration fluid nutrient medium, the each component weight portion of fluid nutrient medium is: peptone 2 g, beef extract 3 g, yeast extract 2 g, manganese sulfate 0.2 g, potassium dihydrogen phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g add water and are settled to 1000mL; PH is controlled at 7.0-7.2; Be sub-packed in the triangular flask of several 500mL, every triangular flask loading amount is 220mL; The rear 30min that sterilizes at 121 ℃ of temperature of sealing, cooling rear standby;
Under aseptic condition, the slant strains that fluid nutrient medium in each triangular flask access step (2) makes, wherein the bacillus licheniformis inoculum concentration be in triangular flask the mixture total weight amount 10%, triangular flask is carried out shaking table cultivates, temperature is controlled at 37 ℃, and rotating speed is 180-200 r/min; Shaking table makes the triangular flask liquid spawn after cultivating 24h, and is standby.
(4) Cans solid fermentation
By formula requirement configuration solid fermentation culture medium, the each component weight portion of solid fermentation culture medium is: brown sugar 0.5%, lime 0.5%, and sterilized water 45%, all the other are wheat bran;
Be distributed in some Cans after mixing the solid fermentation culture medium evenly, bottleneck covers with newspaper, and is tight with linear system; At the temperature of 121 ℃ to above-mentioned Cans sterilization 60 min; After cooling, the liquid spawn that access step (3) is produced, wherein the bacillus licheniformis inoculum concentration is 5% of mixture total weight amount; Put into and cultivate 60 h between the cultivation of 37 ℃, when spore forming rate reaches 90% above fermentation ends, make finished product after discharging.
Further, the sealing in described step (2) refers to cover tampon on test tube, and tampon and test tube notch portion are wrapped newspaper, and is tight with linear system.
Further, in step (1) in every test tube the loading amount of slant medium be the 1/3-1/2 of test tube total capacity.
Further, also comprise the step that the finished product that will make further ferments in step (4) in the fermentation porcelain dish, fermentation temperature is controlled at 37 ℃, and incubation time is 60 h.Described fermentation porcelain dish is common porcelain dish, further ferments at the fermentation porcelain dish, plays to enlarge the effect of cultivating, and can improve output.
The invention has the beneficial effects as follows: mainly utilize the medium of this cheapness of wheat bran as main culture substrate, effectively reduce production cost.Domestic many scholars study the fermentation condition of bacillus licheniformis, but used medium is laboratory glucose commonly used, yeast extract, urea etc., and the industrial production cost is very high.
Adopt solid fermentation method, solid fermentation to have water saving, energy-conservation unique advantage, production cost is low, high temperature resistant, acid and alkali-resistance, anti-extruding, impaired little in the feed process, addition is little, feeding effect is more obvious, belong to clearer production technology, have the potential of large-scale operation.
Adopt this cheap round of Cans, equipment investment is low.Because the fermenting space of each Cans is separate, be convenient to carry out the substrate Characteristics Detection, be convenient to check ferment effect, be convenient to test and controlled fermentation parameter, be convenient to use single factor and orthogonal test of multiple factors that the fermentation condition of bacillus licheniformis is optimized, thereby the optimum temperature of obtaining and pH are convenient to analyze inoculum concentration to the impact of fermentation period; Can formulate flexibly the countermeasure of avoiding microbiological contamination, be convenient to water activity, ventilation and mass transfer, heat transfer and pH etc. are controlled; Miscellaneous bacteria in each round is not easy to influence each other.Environment is relatively controlled, and rocks evenly, and temperature is even.Under this subenvironment of Cans, bacterial classification is by natural selection, and the bacterial strain activity of generation is high, is convenient to enlarge cultivate.
Ferment effect is good.Adopt this technique, the solid fermentation viable count of bacillus licheniformis reaches 5.0 * 10
10Cfu/g has improved bacillus licheniformis output effectively.Keep the viable bacteria rate〉90%, keep about 85% bioactive ingredients.Sanitary index meets additive for microbe feedstuff bacillus licheniformis sanitary index.
Production cost is low.Utilize solid fermentation method, the bacillus licheniformis that produces as culture substrate with wheat bran has production cost to be increased less, the characteristics that unit feed addition obviously reduces are added 100 g bacillus licheniformis with complete feed per ton and are calculated, 5 yuan of costs of feed saving per ton.In application facet, improved the production performance of animal, reduce the generation of disease, can the antibiotic consumption of Substitute For Partial, aspect water quality improvement, effectively organic in decomposition water, the content of ammonia nitrogen in reduction water, for aquatic livestock provides good growing environment, for the development of animal husbandry provides a favorable security.
The present invention has adopted unique solid fermentation culture medium and solid-state fermentation technology, has accelerated culture propagation speed, has shortened fermentation period, has reduced production cost, has improved the sporulation rate of product.
The specific embodiment
Be further described below by embodiment.
Embodiment 1.Utilize bacillus licheniformis to prepare feed addictive, comprise the steps:
(1) bacterial classification preparation and activation
The bacterial classification of the bacillus licheniformis of buying is made suspension, and adding sterilized water to be diluted to concentration is 10
-8The time, on the solid plate culture medium, it being coated with and separating or the line separation, the single bacterium colony of picking part carries out the mensuration of production capacity; Through repeated screening, determine the active bacterial strain strong, that viable count is high of the gemma that produces through fermentation and replace original bacterial strain.Bacillus licheniformis strain is buied from Chinese agriculture microorganism fungus kind preservation administrative center.
(2) slant strains is made
By formula requirement configuration slant medium, the each component weight portion of slant medium is: beef extract 5 g, and peptone 10 g, NaCl 5 g, agar powder 20 g add water and are settled to 1000 mL; PH is controlled at 7.2;
Each component is fully dissolved slant medium heating, heating and temperature control mixes up pH at 50-60 ℃, then is sub-packed in some test tubes, and in every test tube, the loading amount of slant medium is 1/3 of test tube total capacity; Cover tampon on test tube, tampon and test tube notch portion are wrapped newspaper, tight with linear system; Test tube in 121 ℃ of sterilization 30 min, is taken out and puts the inclined-plane, cooling rear standby;
Above-mentioned slant medium is placed in drying box or incubator places the bacterial classification that 24 h access steps (1) prepare, wherein the bacillus licheniformis inoculum concentration be in test tube the mixture total weight amount 1%; Cultivate 24 h, bevel bacterial classification under 37 ℃ of conditions.
(3) the triangular flask liquid spawn is made
By formula requirements configuration fluid nutrient medium, the each component weight portion of fluid nutrient medium is: peptone 2 g, beef extract 3 g, yeast extract 2 g, manganese sulfate 0.2 g, potassium dihydrogen phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, add water calmly to 1000 mL; PH is controlled at 7.0; Be sub-packed in the triangular flask of several 500mL, every triangular flask loading amount is 220 mL; Rear 30 min that sterilize at 121 ℃ of temperature of sealing, cooling rear standby;
Under aseptic condition, the slant strains that fluid nutrient medium in each triangular flask access step (2) makes, wherein the bacillus licheniformis inoculum concentration be in triangular flask the mixture total weight amount 10%; Triangular flask is carried out shaking table cultivate, temperature is controlled at 37 ℃, and rotating speed is 180-200 r/min; Shaking table makes the triangular flask liquid spawn after cultivating 24 h, and is standby.
(4) Cans solid fermentation
By formula requirement configuration solid fermentation culture medium, the each component weight portion of solid fermentation culture medium is: brown sugar 0.5%, lime 0.5%, and sterilized water 45%, all the other are wheat bran;
Be distributed in some Cans after mixing the solid fermentation culture medium evenly, bottleneck covers with newspaper, and is tight with linear system; At the temperature of 121 ℃ to above-mentioned Cans sterilization 60 min;
After cooling, the liquid spawn that access step (3) is produced, wherein the bacillus licheniformis inoculum concentration is 5% of gross weight; Put into and cultivate 60 h between the cultivation of 37 ℃, when spore forming rate reaches 90% above fermentation ends, make finished product after discharging.
Embodiment 2.Utilize bacillus licheniformis to prepare feed addictive, comprise the steps:
(1) bacterial classification preparation and activation
The bacterial classification of the bacillus licheniformis of buying is made suspension, and adding sterilized water to be diluted to concentration is 10
-8The time, to its separation of ruling, the single bacterium colony of picking part carries out the mensuration of production capacity on the solid plate culture medium; Through repeated screening, determine the active bacterial strain strong, that viable count is high of the gemma that produces through fermentation and replace original bacterial strain.
(2) slant strains is made
By formula requirement configuration slant medium, the each component weight portion of slant medium is: beef extract 5 g, and peptone 10 g, NaCl 5 g, agar powder 20 g add water constant volume 1000 mL to end; PH is controlled at 7.4;
Each component is fully dissolved slant medium heating, mix up the pH value, then be sub-packed in some test tubes, in every test tube, the loading amount of slant medium is 1/2 of test tube total capacity; Cover tampon on test tube, tampon and test tube notch portion are wrapped newspaper, tight with linear system; Test tube in 121 ℃ of sterilization 30 min, is taken out and puts the inclined-plane, cooling rear standby;
With above-mentioned slant medium be placed in drying box or incubator place vigor that 48h access step (1) screens high bacterial classification, wherein the bacillus licheniformis inoculum concentration be in test tube the mixture total weight amount 1%; Cultivate 24 h, bevel bacterial classification under 37 ℃ of conditions.
(3) the triangular flask liquid spawn is made
By formula requirement configuration fluid nutrient medium, the each component weight portion of fluid nutrient medium is: peptone 2 g, beef extract 3 g, yeast extract 2 g, manganese sulfate 0.2 g, potassium dihydrogen phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, Jia Shui are settled to 1000 mL; PH is controlled at 7.2; Be sub-packed in the triangular flask of several 500 mL, every triangular flask loading amount is 220 ml; Rear 30 min that sterilize at 121 ℃ of temperature of sealing, cooling rear standby;
Under aseptic condition, the slant strains that fluid nutrient medium in each triangular flask access step (2) makes, wherein the bacillus licheniformis inoculum concentration be in triangular flask the mixture total weight amount 10%; Triangular flask is carried out shaking table cultivate, temperature is controlled at 37 ℃, and rotating speed is 180-200 r/min; Shaking table makes the triangular flask liquid spawn after cultivating 24 h, and is standby.
(4) Cans solid fermentation
By formula requirement configuration solid fermentation culture medium, the each component weight portion of solid fermentation culture medium is: brown sugar 0.5%, lime 0.5%, and sterilized water 45%, all the other are wheat bran;
Be distributed in some Cans after mixing the solid fermentation culture medium evenly, bottleneck covers with newspaper, and is tight with linear system; At the temperature of 121 ℃ to above-mentioned Cans sterilization 60 min;
After cooling, the liquid spawn that access step (3) is produced, wherein the bacillus licheniformis inoculum concentration is 5% of gross weight; Put into and cultivate 60 h between the cultivation of 37 ℃, when spore forming rate reaches 90% above fermentation ends, make finished product after discharging.
Produce the feed addictive viable bacteria content through above-mentioned steps high, total viable count is greater than 5.0 * 10 after testing
10Cfu/g also can keep the viable bacteria rate〉90%, keep about 85% bioactive ingredients.That this composite feed additive has is high temperature resistant, the characteristics of acid and alkali-resistance, anti-bile, and has bin stability preferably.
Embodiment 3.Utilize bacillus licheniformis to prepare feed addictive, comprise the steps:
(1) bacterial classification preparation and activation
The bacterial classification of the bacillus licheniformis of buying is made suspension, and adding sterilized water to be diluted to concentration is 10
-8The time, on the solid plate culture medium, it being coated with separation, the single bacterium colony of picking part carries out the mensuration of production capacity; Through repeated screening, determine the active bacterial strain strong, that viable count is high of the gemma that produces through fermentation and replace original bacterial strain.The active bacterial strain strong, that viable count is high of described gemma refers to its output be compared through test manufacture the bacterial strain of selecting.
(2) slant strains is made
By formula requirement configuration slant medium, the each component weight portion of slant medium is: beef extract 5 g, and peptone 10 g, NaCl 5 g, agar powder 20 g add water and are settled to 1000 mL; PH is controlled at 7.2;
Each component is fully dissolved slant medium heating, mix up the pH value, then be sub-packed in some test tubes, in every test tube, the loading amount of slant medium is 1/3 of test tube total capacity; Cover tampon on test tube, tampon and test tube notch portion are wrapped newspaper, tight with linear system; Test tube in 121 ℃ of sterilization 30 min, is taken out and puts the inclined-plane, cooling rear standby;
Above-mentioned slant medium is placed in drying box or incubator places the bacterial classification that 48h access step (1) prepares, in each test tube the bacillus licheniformis inoculum concentration be in test tube the mixture total weight amount 1%; Cultivate 24 h, bevel bacterial classification under 37 ℃ of conditions.
(3) the triangular flask liquid spawn is made
By formula requirement configuration fluid nutrient medium, the each component weight portion of fluid nutrient medium is: peptone 2 g, beef extract 3 g, yeast extract 2 g, manganese sulfate 0.2 g, potassium dihydrogen phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, Jia Shui are settled to 1000 mL; PH is controlled at 7.2; Be sub-packed in the triangular flask of several 500mL, every triangular flask loading amount is 220 ml; Rear 30 min that sterilize at 121 ℃ of temperature of sealing, cooling rear standby;
Under aseptic condition, the slant strains that fluid nutrient medium in each triangular flask access step (2) makes, wherein the bacillus licheniformis inoculum concentration be in triangular flask the mixture total weight amount 10%; Triangular flask is carried out shaking table cultivate, temperature is controlled at 37 ℃, and rotating speed is 180-200 r/min; Shaking table makes the triangular flask liquid spawn after cultivating 24 h, and is standby.
(4) Cans solid fermentation
By formula requirement configuration solid fermentation culture medium, the each component weight portion of solid fermentation culture medium is: brown sugar 0.5%, lime 0.5%, and sterilized water 45%, all the other are wheat bran;
Be distributed in some Cans after mixing the solid fermentation culture medium evenly, bottleneck covers with newspaper, and is tight with linear system; At the temperature of 121 ℃ to above-mentioned Cans sterilization 60 min;
After cooling, the liquid spawn that access step (3) is produced, wherein the bacillus licheniformis inoculum concentration is 5% of gross weight; Put into and cultivate 60 h between the cultivation of 37 ℃, when spore forming rate reaches 90% above fermentation ends, discharging makes finished product;
The finished product that step 4 is made further ferments in the fermentation porcelain dish, and fermentation temperature is controlled at 37 ℃, and incubation time is 60 h.Through further fermentation, improve production.Due to the finite capacity of Cans, the effect that enlarges cultivation has been played in further fermentation in the fermentation porcelain dish.Ferment in the fermentation porcelain dish, be convenient to stir material, make its fermentation evenly.
Produce the feed addictive viable bacteria content through above-mentioned steps high, detect total viable count greater than 5.0 * 10 through feeding quality supervision and inspection center of the Ministry of Agriculture
10Cfu/g keeps the viable bacteria rate〉90%, keep about 85% bioactive ingredients.That this composite feed additive has is high temperature resistant, the characteristics of acid and alkali-resistance, anti-bile, and has bin stability preferably.
Experimental example 1.According in the experiment of Shandong Jin Ming Industrial Co., Ltd., select to differ from and delivered totally 12 of Landraces in two months for sale, be divided into control group and test group, every group each 6.Control group is selected plump large swine feed 305, does not add any other thing feeding.Test group is selected plump large swine feed 305 equally, adds in the ratio of 1000g/t the feed addictive that the present invention makes simultaneously, and experimental period is 2 months.Result is as follows:
0.1% feed addictive of table 1 is to differing from the Landrace weight gain situation of delivering for sale in two months
| ? | The test initial weight | The test end is heavy | Weightening finish altogether | Average daily gain |
| Control group | 561 | 1194 | 633 | 9.59 |
| Experimental group | 570 | 1272 | 702 | 10.64 |
0.1% feed addictive of table 2 is to differing from the Landrace feed conversion rate situation of delivering for sale in two months
| ? | Consume feed | Feedstuff-meat ratio |
| Control group | 1920 | 3.033 |
| Experimental group | 2000 | 2.849 |
Table 3 economic benefit affects situation
Experimental example 2.Be the effect of feed addictive in the Commercial meat-type duck large-scale cultivation that checking the present invention makes, illustrious and influential agricultural development Co., Ltd tests in Shandong.Three pilot regions have been selected in the Gaotang County, Shandong Province, 4 of the Cherry Village Ducks plants that each zone selects respectively that 4 cultivation scales approach, cultivating condition is basically identical, be divided at random 2 groups of each 2 plants, use respectively basal diet and add the test daily ration of the feed addictive that the present invention makes, meat duck 44 ages in days are delivered for sale.In this test, plant's basal diet used and test daily ration provide by Shandong six directions feed corporation,Ltd, and basal diet is prepared according to U.S. NRC (1994) meat duck feeding standard.In this research, plant is all according to requiring feeding and management meat duck in daily production, and provides application technology to instruct by the technical staff, and in same pilot region, the management of plant is consistent as far as possible.In test, no matter control group and test group all according to duck public sentiment condition standardized drug, forbid blindly making medicine.The indexs such as the full phase feed consumption of test of statistics meat duck survival rate, every plumage duck, the trunk weightening finish of full phase of test, feedstuff-meat ratio and average expenses for medicine cost of full phase in test.Concrete condition sees Table 4.
Situation and the daily ration arrangement of table 4 test duck field
| Pilot region | Cultivation scale (plumage/family) * | The control group daily ration | Test group daily ration bacillus consumption (g/t) |
| One trial zone, family, Gaotang | 3000 plumages | Basal diet | 500 |
| Two trial zones, family, Gaotang | 5000 plumages | Basal diet | 1000 |
| Three trial zones, family, Gaotang | 8000 plumages | Basal diet | 1000 |
It is that meat duck 4 plants of 3000 plumages have participated in test that the cultivation scale has been selected at one family, Gaotang, has wherein added the feed addictive that 100g the present invention makes in test group daily ration per ton, and result of the test sees Table 5.
Add the feed addictive that the present invention makes in one trial zone, the family daily ration of table 5 Gaotang
Impact on meat duck production performance
Annotate: " indicator difference " value is the difference (absolute value) of this row test group index and control group index, and is lower same.
It is that meat duck 4 plants of 5000 plumages have participated in test that the cultivation scale has been selected at two families, Gaotang, considers that this district plant cultivation density is higher, therefore add the feed addictive that 150g the present invention makes in test group daily ration per ton, result of the test sees Table 6.Found by result of the test, the effect of the bacillus licheniformis of this trial zone is consistent with the effect of one trial zone, family, Gaotang, and under the experimental condition of this trial zone, after increasing the bacillus consumption, the amplitude of expenses for medicine cost is larger, test group relative comparison group absolute value reduces by 0.16 yuan/plumage, and relative value reduces by 22.2%.
Add the feed addictive that the present invention makes in two trial zone, the family daily rations of table 6 Gaotang
Impact on meat duck production performance
It is that meat duck 4 plants of 8000 plumages have participated in test that the cultivation scale has been selected at three families, Gaotang, considers that equally this district plant cultivation density is higher, larger, therefore also added the 180g bacillus licheniformis in test group daily ration per ton, result of the test sees Table 7.
Add the feed addictive that the present invention makes in three trial zone, the family daily rations of table 7 Gaotang
Impact on meat duck production performance
The result of the test of having verified front two phases of this district's culture experiment has confirmed that further the feed addictive that the present invention makes improves the effect that Cherry Village Ducks cultivates achievement dry straightly, and the result of use that the feed addictive that the present invention makes is described is very stable.In the test of one's respective area, the reduction of expenses for medicine cost is very outstanding, and test group relative comparison group absolute value reduces by 0.30 yuan/plumage, and relative value reduces and reaches 40.0%.
Comprehensive trizonal culture experiment interpretation of result, the effect of the feed addictive that the present invention makes aspect reduction meat duck cultivation feedstuff-meat ratio is very stable, can reduce feedstuff-meat ratio 0.06～0.07, and the reduction amplitude is between 2.4%～2.7%; And on the aspect that affects of expenses for medicine cost, Different adding amount, different tests condition change greatly, and the every plumage of expenses for medicine reduces 0.08-0.30 unit, and the reduction amplitude is between 11.4%-40.0%.This prompting can be adjusted the bacillus licheniformis consumption in conjunction with the concrete cultivating condition of plant in application in practice, to reach best culture efficiency.In addition, in trizonal test, in the whole breeding process of test group plant, the ight soil drying and moulding is all good than control group, breeding environment improves obviously, bedding and padding are changed number of times and are reduced, this and bacillus licheniformis can be improved the animal intestinal microbial environment inner link, is also a basis that reduces cultivation expenses for medicine cost in production practices.
Two trial zones, family are example take the Gaotang, and brief analysis is heavily used the economic benefit of the feed addictive that the present invention makes at meat duck material.According to table 6 statistics, test group and control group feedstuff-meat ratio improve 2.4%, in trunk unit price 7.44 (hair duck 6.4) unit/kilogram, by the European productivity effect mode computation of considering the factors such as climate change, actual growth by 2.0%, every plumage duck weight increases by 0.06 kilogram, and every plumage duck increases income 0.45 yuan; Simultaneously, every plumage reduces by 0.30 yuan of drug cost; Basal diet calculates by 2.17 yuan/kg kilogram, and the every plumage of test group is saved 0.04 yuan of cost of basal diet (the gemma cost calculates separately); 20 yuan/tons of the feedstuff additive product costs that input the present invention makes, namely 0.02 yuan/kilogram, every plumage meat duck input cost is 0.15 yuan.Thereby every plumage meat duck net output value added is 0.64 yuan, and input-output ratio is 1:7.Calculate like this, delivering 5000 net incomes for sale increases by 3000 yuan, and economic benefit is considerable.
Claims (4)
1. a method of utilizing bacillus licheniformis to produce feed addictive, is characterized in that comprising the steps:
(1) bacterial classification preparation and activation
The bacterial classification of the bacillus licheniformis of buying is made suspension, and adding sterilized water to be diluted to concentration is 10
-8The time, on the solid plate culture medium, it being coated with and separating or the line separation, the single bacterium colony of picking part carries out the mensuration of production capacity; Through repeated screening, determine the active bacterial strain strong, that viable count is high of the gemma that produces through fermentation and replace original bacterial strain;
(2) slant strains is made
By formula requirement configuration slant medium, the each component weight portion of slant medium is: beef extract 5 g, and peptone 10 g, NaCl 5 g, agar powder 20-25 g adds water and is settled to 1000 mL; PH is controlled at 7.2-7.4;
Each component is fully dissolved the slant medium heating, mix up pH, then be sub-packed in some test tubes, the sealing test tube; Test tube in 121 ℃ of sterilization 30 min, is taken out and puts the inclined-plane, cooling rear standby;
Above-mentioned slant medium is placed in drying box or incubator places the bacterial classification that 24-48h access step (1) prepares, wherein the bacillus licheniformis inoculum concentration be in test tube the mixture total weight amount 1%, cultivate 24h under 37 ℃ of conditions, the bevel bacterial classification;
(3) the triangular flask liquid spawn is made
By formula requirement configuration fluid nutrient medium, the each component weight portion of fluid nutrient medium is: peptone 2 g, beef extract 3 g, yeast extract 2 g, manganese sulfate 0.2 g, potassium dihydrogen phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g add water and are settled to 1000mL; PH is controlled at 7.0-7.2; Be sub-packed in the triangular flask of several 500mL, every triangular flask loading amount is 220mL; The rear 30min that sterilizes at 121 ℃ of temperature of sealing, cooling rear standby;
Under aseptic condition, the slant strains that fluid nutrient medium in each triangular flask access step (2) makes, wherein the bacillus licheniformis inoculum concentration be in triangular flask the mixture total weight amount 10%, triangular flask is carried out shaking table cultivates, temperature is controlled at 37 ℃, and rotating speed is 180-200 r/min; Shaking table makes the triangular flask liquid spawn after cultivating 24h, and is standby;
(4) Cans solid fermentation
By formula requirement configuration solid fermentation culture medium, the each component weight portion of solid fermentation culture medium is: brown sugar 0.5%, lime 0.5%, and sterilized water 45%, all the other are wheat bran;
Be distributed in some Cans after mixing the solid fermentation culture medium evenly, bottleneck covers with newspaper, and is tight with linear system; At the temperature of 121 ℃ to above-mentioned Cans sterilization 60 min;
After cooling, the liquid spawn that access step (3) is produced, wherein the bacillus licheniformis inoculum concentration is 5% of mixture total weight amount; Put into and cultivate 60 h between the cultivation of 37 ℃, when spore forming rate reaches 90% above fermentation ends, make finished product after discharging.
2. a kind of method of utilizing bacillus licheniformis to produce feed addictive as claimed in claim 1, it is characterized in that: the sealing in described step (2) refers to cover tampon on test tube, and tampon and test tube notch portion are wrapped newspaper, and is tight with linear system.
3. a kind of method of utilizing bacillus licheniformis to produce feed addictive as claimed in claim 1 is characterized in that: in every test tube, the loading amount of slant medium is the 1/3-1/2 of test tube total capacity in step (2).
4. a kind of described method of utilizing bacillus licheniformis to produce feed addictive of a claim as any in claim 1 to 3, it is characterized in that: also comprise the step that the finished product that will make further ferments in step (4) in the fermentation porcelain dish, fermentation temperature is controlled at 37 ℃, and incubation time is 60 h.
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Denomination of invention: A method of using Bacillus licheniformis to produce feed additives Effective date of registration: 20231130 Granted publication date: 20150121 Pledgee: Shandong Gaotang Rural Commercial Bank Co.,Ltd. Pledgor: GAOTANG HUANONG BIOENGINEERING CO.,LTD. Registration number: Y2023980068527 |