CN103109981B - 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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Landscapes
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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 fodder additives.
Background technology
Raiser cultivate domestic birds and animals time in order to reach prophylactic object, often in feed, add a large amount of microbiotic, consequently inevitably cause abuse of antibiotics, produce drug residue in animal body, accumulate in human body eventually through food chain, serious threat human health.Research shows that Bacillus licheniformis is all class probiotic bacteriums, add in feed can function as follows with the preparation that it is made: 1. genus bacillus is a kind of aerobic probiotics, it can form spore in adverse environment, and high temperature resistant, acid and alkali-resistance and temperature variation, can bring back to life rapidly after entering enteron aisle; And by fighting for limited nutritive substance with pathogenic bacterium, consume limited oxygen in environment rapidly, be conducive to maintaining intestinal flora balance and play an important role, thus suppress enteropathic bacteria growing, reach the effect of prophylactic treatment livestock and poultry intestinal canal diseases; 2. genus bacillus can produce multiple digestive ferment, helps animal digesting and assimilating nutritive substance, thus promoting digestion and raising efficiency of feed utilization, reduce feeding cost; 3. genus bacillus can promote that animal intestinal associated lymphoid tissue is in " the immune ready state " of height, makes Development of Immune Organs accelerate simultaneously, and immune system maturation is morning soon, T, bone-marrow-derived lymphocyte increasing number, and animal body fluid and cellular immune level improve; 4. genus bacillus growth and breeding in animal intestinal, can produce multiple nutrients material as VITAMIN, amino acid, organic acid, somatomedin etc., participates in animal body metabolism, for body provides nutritive substance.
Because bacillus preparation has above-mentioned important excellent characteristic, therefore numerous Chinese scholars has been attracted to further investigate it, research contents mainly concentrates on: the screening (Yang Xiaobin of genus bacillus, Xie Yongkui, Wei Yanpeng etc., the screening of Bacillus Subtilis Natto of Probiotics, Guangzhou Food Industry science and technology, supplementary issue in 2003), (complementary work is protected to prepare plasmin, Guo Ailing, Qin Qiaoling etc., the screening of one strain high-yield nattokinase bacterial strain and the optimization of fermentation condition thereof, Food science, 4th phase in 2007), deep liquid fermentation process (Zhu Jianhui, Du Lianxiang, Lu Fuping, Nattokinase liquid-state fermentation technology is optimized, food and fermentation industries, 8th phase in 2005), pulvis preparation technology (Zhong Qingping, Wang Faxiang, Liu Jiachang, the development of Bacillus natto viable bacteria drying agent, foodstuffs industry science and technology, 8th phase in 2005) etc. aspect, and concrete solid fermentation process has no report.
The tolerances such as solid fermentation method compares that the maximum advantage of liquid fermenting is exactly that production cost is low, starting material are simple, product inulinase-producing activity is high, high temperature resistant, acid and alkali-resistance, the anti-extrusion of product are strong.But solid-fermented technique difficulty is large, is not easy to produce high-quality bacillus preparation.The preparation viable bacteria content that the production technique of current bacillus preparation and fermentative medium formula are produced is not high, and preparation validity period is short, makes product be difficult to reach desirable disease prevention and growth-promoting effect.
Summary of the invention
Content of the present invention is, not enough for prior art, provides a kind of solid fermentation cultural method utilizing Bacillus licheniformis to prepare fodder additives, the fodder additives viable bacteria content that the method obtains is high, and miscellaneous bacteria rate is low, and brood cell is active strong, stability is strong, and production cost is not high.
The technical solution used in the present invention is, a kind of method utilizing Bacillus licheniformis to produce fodder additives, 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 be 10
-8time, solid plate substratum carries out coating to it and is separated or line separation, picking part list bacterium colony carries out the mensuration of throughput; Through repeated screening, determine that active strong, that viable count the is high bacterial strain of the gemma produced through fermentation replaces original bacterial strain.
(2) slant strains makes
By recipe requirements configuration slant medium, each component weight part of slant medium is: extractum carnis 5 g, peptone 10 g, NaCl 5 g, and agar powder 20-25 g, adds water and be settled to 1000 mL; PH controls at 7.2-7.4;
Each component is fully dissolved slant medium heating, mix up pH, be then sub-packed in some test tubes, sealing test tube; By test tube in 121 DEG C of sterilizing 30 min, take out pendulum inclined-plane, for subsequent use after cooling;
Above-mentioned slant medium is placed in loft drier or incubator to place 24-48h and access bacterial classification prepared by step (1), wherein Bacillus licheniformis inoculum size is 1% of mixture total weight amount in test tube, under 37 DEG C of conditions, cultivate 24h, bevel bacterial classification.
(3) triangular flask liquid spawn makes
By recipe requirements configuration liquid nutrient medium, the each component weight part of liquid nutrient medium is: peptone 2 g, extractum carnis 3 g, yeast extract paste 2 g, manganous sulfate 0.2 g, potassium primary phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, add water and be settled to 1000mL; PH controls at 7.0-7.2; Be sub-packed in the triangular flask of several 500mL, often propping up triangular flask loading amount is 220mL; After sealing at 121 DEG C of temperature sterilizing 30min, cooling after for subsequent use;
Aseptically, the slant strains that liquid nutrient medium access step (2) in each triangular flask is obtained, wherein Bacillus licheniformis inoculum size is 10% of mixture total weight amount in triangular flask, carries out shaking table cultivation to triangular flask, temperature controls at 37 DEG C, and rotating speed is 180-200 r/min; Shaking table is obtained triangular flask liquid spawn after cultivating 24h, for subsequent use.
(4) Cans solid fermentation
By recipe requirements configuration solid fermentation substratum, each component weight part of solid fermentation substratum is: brown sugar 0.5%, lime 0.5%, sterilized water 45%, and all the other are wheat bran;
Be distributed into after being mixed evenly by solid fermentation substratum in some Cans, bottleneck newspaper covers, tight by linear system; To above-mentioned Cans sterilizing 60 min at the temperature of 121 DEG C; After cooling, the liquid spawn that access step (3) is produced, wherein Bacillus licheniformis inoculum size is 5% of mixture total weight amount; 60 h are cultivated between the cultivation of putting into 37 DEG C, when spore forming rate reaches more than 90% fermentation ends, discharging.
Further, the sealing in described step (2) refers to and cover tampon on test tube, and tampon and test tube notch portion are wrapped newspaper, tight by linear system.
Further, the loading amount of often propping up slant medium in test tube is in step (1) the 1/3-1/2 of test tube total volume.
Further, in step (4), also comprise the step of being fermented in fermentation porcelain dish further by obtained material, leavening temperature controls at 37 DEG C, and incubation time is 60 h.Described fermentation porcelain dish is common porcelain dish, ferments further, play the effect of enlarged culturing, can improve output at fermentation porcelain dish.
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.The fermentation condition of domestic many scholars to Bacillus licheniformis is studied, but used medium is glucose, yeast extract, urea etc. that laboratory is commonly used, and industrial production cost is very high.
Adopt solid fermentation method, solid fermentation has water saving, energy-conservation unique advantage, and production cost is low, high temperature resistant, acid and alkali-resistance, anti-extrusion, impaired little in feed manufacturing process, addition is little, feeding effect is more obvious, belong to clearer production technology, there is the potential of large-scale operation.
Adopt this cheap fermenting container of Cans, facility investment is low.Because the fermenting space of each Cans is separate, be convenient to carry out substrate specificities detection, be convenient to check ferment effect, be convenient to test and control fermentation parameter, be convenient to use single factor test and the fermentation condition of orthogonal test of multiple factors to Bacillus licheniformis to be optimized, thus obtain optimum temps and pH, be convenient to analyze inoculum size to the impact of fermentation period; The countermeasure avoiding microbiological contamination can be formulated flexibly, be convenient to control water activity, ventilation and mass transfer, heat transfer and pH etc.; Miscellaneous bacteria in each fermenting container is not easy to influence each other.Environment compares control, rocks evenly, homogeneous temperature.Under this subenvironment of Cans, bacterial classification is by natural selection, and the strain activity of generation is high, is convenient to enlarged culturing.
Ferment effect is good.Adopt this technique, the solid fermentation viable count of Bacillus licheniformis reaches 5.0 × 10
10cfu/g, improves Bacillus licheniformis output effectively.Keep viable bacteria rate >90%, retain about 85% bioactive ingredients.Sanitary index meets microorganism feed addictive Bacillus licheniformis sanitary index.
Production cost is low.Utilize solid fermentation method, the Bacillus licheniformis produced using wheat bran as culture substrate has production cost to be increased less, the feature that unit feed addition obviously reduces, adds 100 g Bacillus licheniformis with complete feed per ton and calculates, feed saving per ton 5 yuan of costs.In application aspect, improve the production performance of animal, reduce the generation of disease, can the antibiotic consumption of Substitute For Partial, in water quality improvement, organic in effective water of decomposition, reduce the content of ammonia nitrogen in water, for aquatic animal provides good growing environment, for the development of livestock industry provides a favorable security.
Present invention employs unique solid fermentation substratum and solid-state fermentation technology, accelerate culture propagation speed, shorten fermentation period, reduce production cost, improve the sporulation rate of product.
Embodiment
Be further described below by embodiment.
Embodiment 1.Utilize Bacillus licheniformis to prepare fodder additives, 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 be 10
-8time, solid plate substratum carries out coating to it and is separated or line separation, picking part list bacterium colony carries out the mensuration of throughput; Through repeated screening, determine that active strong, that viable count the is high bacterial strain of the gemma produced through fermentation replaces original bacterial strain.Bacillus licheniformis strain is buied from Chinese agriculture Microbiological Culture Collection administrative center.
(2) slant strains makes
By recipe requirements configuration slant medium, each component weight part of slant medium is: extractum carnis 5 g, peptone 10 g, NaCl 5 g, and agar powder 20 g, adds water and be settled to 1000 mL; PH controls 7.2;
Each component is fully dissolved slant medium heating, heating and temperature control, at 50-60 DEG C, mixes up pH, is then sub-packed in some test tubes, and the loading amount of often propping up slant medium in test tube is 1/3 of test tube total volume; Test tube covers tampon, tampon and test tube notch portion are wrapped newspaper, tight by linear system; By test tube in 121 DEG C of sterilizing 30 min, take out pendulum inclined-plane, for subsequent use after cooling;
Above-mentioned slant medium is placed in loft drier or incubator to place 24 h and access bacterial classification prepared by step (1), wherein Bacillus licheniformis inoculum size is 1% of mixture total weight amount in test tube; 24 h are cultivated, bevel bacterial classification under 37 DEG C of conditions.
(3) triangular flask liquid spawn makes
By recipe requirements configuration liquid nutrient medium, each component weight part of liquid nutrient medium is: peptone 2 g, extractum carnis 3 g, yeast extract paste 2 g, manganous sulfate 0.2 g, potassium primary phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, add water fixed to 1000 mL; PH controls 7.0; Be sub-packed in the triangular flask of several 500mL, often propping up triangular flask loading amount is 220 mL; After sealing at 121 DEG C of temperature sterilizing 30 min, cooling after for subsequent use;
Aseptically, the slant strains that liquid nutrient medium access step (2) in each triangular flask is obtained, wherein Bacillus licheniformis inoculum size is 10% of mixture total weight amount in triangular flask; Carry out shaking table cultivation to triangular flask, temperature controls at 37 DEG C, and rotating speed is 180-200 r/min; Shaking table is obtained triangular flask liquid spawn after cultivating 24 h, for subsequent use.
(4) Cans solid fermentation
By recipe requirements configuration solid fermentation substratum, each component weight part of solid fermentation substratum is: brown sugar 0.5%, lime 0.5%, sterilized water 45%, and all the other are wheat bran;
Be distributed into after being mixed evenly by solid fermentation substratum in some Cans, bottleneck newspaper covers, tight by linear system; To above-mentioned Cans sterilizing 60 min at the temperature of 121 DEG C;
After cooling, the liquid spawn that access step (3) is produced, wherein Bacillus licheniformis inoculum size is 5% of gross weight; 60 h are cultivated between the cultivation of putting into 37 DEG C, when spore forming rate reaches more than 90% fermentation ends, obtained finished product after discharging.
Embodiment 2.Utilize Bacillus licheniformis to prepare fodder additives, 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 be 10
-8time, solid plate substratum carries out line to it and is separated, picking part list bacterium colony carries out the mensuration of throughput; Through repeated screening, determine that active strong, that viable count the is high bacterial strain of the gemma produced through fermentation replaces original bacterial strain.
(2) slant strains makes
By recipe requirements configuration slant medium, each component weight part of slant medium is: extractum carnis 5 g, peptone 10 g, NaCl 5 g, agar powder 20 g, and add water constant volume 1000 mL to end; PH controls 7.4;
Each component is fully dissolved slant medium heating, mix up pH value, be then sub-packed in some test tubes, the loading amount of often propping up slant medium in test tube is 1/2 of test tube total volume; Test tube covers tampon, tampon and test tube notch portion are wrapped newspaper, tight by linear system; By test tube in 121 DEG C of sterilizing 30 min, take out pendulum inclined-plane, for subsequent use after cooling;
Above-mentioned slant medium is placed in loft drier or incubator place 48h access vigor that step (1) screens high bacterial classification, wherein Bacillus licheniformis inoculum size is 1% of mixture total weight amount in test tube; 24 h are cultivated, bevel bacterial classification under 37 DEG C of conditions.
(3) triangular flask liquid spawn makes
By recipe requirements configuration liquid nutrient medium, each component weight part of liquid nutrient medium is: peptone 2 g, extractum carnis 3 g, yeast extract paste 2 g, manganous sulfate 0.2 g, potassium primary phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, adding water is settled to 1000 mL; PH controls 7.2; Be sub-packed in the triangular flask of several 500 mL, often propping up triangular flask loading amount is 220 ml; After sealing at 121 DEG C of temperature sterilizing 30 min, cooling after for subsequent use;
Aseptically, the slant strains that liquid nutrient medium access step (2) in each triangular flask is obtained, wherein Bacillus licheniformis inoculum size is 10% of mixture total weight amount in triangular flask; Carry out shaking table cultivation to triangular flask, temperature controls at 37 DEG C, and rotating speed is 180-200 r/min; Shaking table is obtained triangular flask liquid spawn after cultivating 24 h, for subsequent use.
(4) Cans solid fermentation
By recipe requirements configuration solid fermentation substratum, each component weight part of solid fermentation substratum is: brown sugar 0.5%, lime 0.5%, sterilized water 45%, and all the other are wheat bran;
Be distributed into after being mixed evenly by solid fermentation substratum in some Cans, bottleneck newspaper covers, tight by linear system; To above-mentioned Cans sterilizing 60 min at the temperature of 121 DEG C;
After cooling, the liquid spawn that access step (3) is produced, wherein Bacillus licheniformis inoculum size is 5% of gross weight; 60 h are cultivated between the cultivation of putting into 37 DEG C, when spore forming rate reaches more than 90% fermentation ends, obtained finished product after discharging.
Produce fodder additives viable bacteria content through above-mentioned steps high, total viable count is greater than 5.0 × 10 after testing
10cfu/g, also can keep viable bacteria rate >90%, retains about 85% bioactive ingredients.This composite feed additive has high temperature resistant, acid and alkali-resistance, resistance to bile feature, and has good package stability.
Embodiment 3.Utilize Bacillus licheniformis to prepare fodder additives, 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 be 10
-8time, solid plate substratum carries out coating to it and is separated, picking part list bacterium colony carries out the mensuration of throughput; Through repeated screening, determine that active strong, that viable count the is high bacterial strain of the gemma produced through fermentation replaces original bacterial strain.Active strong, that viable count the is high bacterial strain of described gemma refers to through test manufacture, compares, the bacterial strain selected to its output.
(2) slant strains makes
By recipe requirements configuration slant medium, each component weight part of slant medium is: extractum carnis 5 g, peptone 10 g, NaCl 5 g, and agar powder 20 g, adds water and be settled to 1000 mL; PH controls 7.2;
Each component is fully dissolved slant medium heating, mix up pH value, be then sub-packed in some test tubes, the loading amount of often propping up slant medium in test tube is 1/3 of test tube total volume; Test tube covers tampon, tampon and test tube notch portion are wrapped newspaper, tight by linear system; By test tube in 121 DEG C of sterilizing 30 min, take out pendulum inclined-plane, for subsequent use after cooling;
Above-mentioned slant medium is placed in loft drier or incubator to place 48h and access bacterial classification prepared by step (1), in each test tube, Bacillus licheniformis inoculum size is 1% of mixture total weight amount in test tube; 24 h are cultivated, bevel bacterial classification under 37 DEG C of conditions.
(3) triangular flask liquid spawn makes
By recipe requirements configuration liquid nutrient medium, each component weight part of liquid nutrient medium is: peptone 2 g, extractum carnis 3 g, yeast extract paste 2 g, manganous sulfate 0.2 g, potassium primary phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, adding water is settled to 1000 mL; PH controls 7.2; Be sub-packed in the triangular flask of several 500mL, often propping up triangular flask loading amount is 220 ml; After sealing at 121 DEG C of temperature sterilizing 30 min, cooling after for subsequent use;
Aseptically, the slant strains that liquid nutrient medium access step (2) in each triangular flask is obtained, wherein Bacillus licheniformis inoculum size is 10% of mixture total weight amount in triangular flask; Carry out shaking table cultivation to triangular flask, temperature controls at 37 DEG C, and rotating speed is 180-200 r/min; Shaking table is obtained triangular flask liquid spawn after cultivating 24 h, for subsequent use.
(4) Cans solid fermentation
By recipe requirements configuration solid fermentation substratum, each component weight part of solid fermentation substratum is: brown sugar 0.5%, lime 0.5%, sterilized water 45%, and all the other are wheat bran;
Be distributed into after being mixed evenly by solid fermentation substratum in some Cans, bottleneck newspaper covers, tight by linear system; To above-mentioned Cans sterilizing 60 min at the temperature of 121 DEG C;
After cooling, the liquid spawn that access step (3) is produced, wherein Bacillus licheniformis inoculum size is 5% of gross weight; Cultivate 60 h between the cultivation of putting into 37 DEG C, when spore forming rate reaches more than 90% fermentation ends, discharging obtains finished product;
Fermented in fermentation porcelain dish further by finished product obtained for step 4, leavening temperature controls at 37 DEG C, and incubation time is 60 h.Through fermentation further, improve production.Due to the finite capacity of Cans, ferment further in fermentation porcelain dish, serve the effect of enlarged culturing.Ferment in fermentation porcelain dish, be convenient to stir material, make it ferment evenly.
Produce fodder additives viable bacteria content through above-mentioned steps high, detect total viable count through feeding quality supervision and inspection center of the Ministry of Agriculture and be greater than 5.0 × 10
10cfu/g, keeps viable bacteria rate >90%, retains about 85% bioactive ingredients.This composite feed additive has high temperature resistant, acid and alkali-resistance, resistance to bile feature, and has good package stability.
Experimental example 1.Test according in Shandong Jin Ming Industrial Co., Ltd., choosing differs from two months and delivers landrace totally 12 for sale, is divided into control group and test group, often organizes each 6.Control group selects plump large swine feed 305, does not add any other thing feeding.Test group selects plump large swine feed 305 equally, and add in the ratio of 1000g/t the fodder additives that the present invention obtains, trial period is 2 months simultaneously.Result is as follows:
Table 1 0.1% fodder additives delivers landrace weight gain situation for sale to differing from two months
| ? | Test initial weight | Test end is heavy | Weightening finish altogether | Average daily gain |
| Control group | 561 | 1194 | 633 | 9.59 |
| Experimental group | 570 | 1272 | 702 | 10.64 |
Table 2 0.1% fodder additives delivers landrace feed conversion rate situation for sale to differing from 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.For the effect of fodder additives in Commercial meat-type duck large-scale cultivation that checking the present invention obtains, in Shandong, illustrious and influential agricultural development company limited tests.Have selected three pilot regions in Gaotang County, Shandong Province, 4 Cherry Village Ducks plants 4 that cultivation scale is close, cultivating condition is basically identical are selected in each region respectively, be divided into 2 groups of each 2 plants at random, the test daily ration of the fodder additives using basal diet and interpolation the present invention to obtain respectively, meat duck 44 age in days is 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 according to U.S. NRC (1994) boiler duck feeding standard preparation.In this research, plant all according to the requirement feeding and management meat duck in daily production, and provides utilisation technology to instruct by technician, 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.Add up the test full phase feed consumption of meat duck surviving rate, often plumage duck in test, test the indexs such as full phase trunk weightening finish, feedstuff-meat ratio and full phase average expenses for medicine cost.Particular case is in table 4.
Table 4 tests situation and the daily ration arrangement of duck field
| Pilot region | Cultivation scale (plumage/family) * | Control group daily ration | Test group daily ration genus bacillus consumption (g/t) |
| Test site, family, Gaotang one | 3000 plumages | Basal diet | 500 |
| Test site, family, Gaotang two | 5000 plumages | Basal diet | 1000 |
| Test site, family, Gaotang three | 8000 plumages | Basal diet | 1000 |
It is that meat duck 4 plants of 3000 plumages take part in test that family, Gaotang one have selected cultivation scale, and wherein with the addition of the fodder additives that 100g the present invention obtains in test group daily ration per ton, test-results is in table 5.
The fodder additives that the present invention obtains is added in test site, family, table 5 Gaotang one daily ration
On the impact of meat duck production performance
Note: " indicator difference " value is the difference (absolute value) of this row test group index and control group index, lower same.
It is that meat duck 4 plants of 5000 plumages take part in test that family, Gaotang two have selected cultivation scale, considers that this plant of district cultivation density is higher, therefore adds the fodder additives that 150g the present invention obtains in test group daily ration per ton, and test-results is in table 6.Found by test-results, the effect of the Bacillus licheniformis of this test site is consistent with the effect of test site, family, Gaotang one, and under the test conditions of this test site, after increasing genus bacillus consumption, the amplitude that expenses for medicine cost reduces is larger, test group relative comparison group absolute value reduces by 0.16 yuan/plumage, and relative value then reduces by 22.2%.
The fodder additives that the present invention obtains is added in test site, family, table 6 Gaotang two daily ration
On the impact of meat duck production performance
It is that meat duck 4 plants of 8000 plumages take part in test that family, Gaotang three have selected cultivation scale, and consider that this plant of district cultivation density is higher, larger equally, therefore also with the addition of 180g Bacillus licheniformis in test group daily ration per ton, test-results is in table 7.
The fodder additives that the present invention obtains is added in test site, family, table 7 Gaotang three daily ration
On the impact of meat duck production performance
The test-results demonstrating front two phases of this district's culture experiment, further demonstrate that the fodder additives that the present invention obtains improves the effect that Cherry Village Ducks cultivates achievement, illustrates that the result of use of the fodder additives that the present invention obtains is very stable dry straightly.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 then reduces and reaches 40.0%.
Comprehensive trizonal culture experiment interpretation of result, the effect of the fodder additives that the present invention obtains in reduction meat duck cultivation feedstuff-meat ratio is very stable, and can reduce feedstuff-meat ratio 0.06 ~ 0.07, the amplitude that reduces is between 2.4% ~ 2.7%; And the aspect that affects on 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 amplitude that reduces is between 11.4%-40.0%.This prompting, in practical application, in conjunction with the concrete cultivating condition of plant, can adjust Bacillus licheniformis consumption, to reach best culture efficiency.In addition, in trizonal test, in the whole breeding process of test group plant, all comparatively control group is good for ight soil drying and moulding, breeding environment improves obviously, bedding and padding are changed number of times and are reduced, this and Bacillus licheniformis can improve animal intestinal microbial environment inner link, is also the basis reducing cultivation expenses for medicine cost in production practice.
For test site, family, Gaotang two, brief analysis heavily applies the economic benefit of the fodder additives that the present invention obtains 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 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; Meanwhile, every plumage reduces drug cost 0.30 yuan; Basal diet calculates by 2.17 yuan/kg kilogram, then the every plumage of test group saves basal diet cost 0.04 yuan (gemma cost calculates separately); Drop into the feedstuff additive product cost 20 yuan/ton that the present invention obtains, namely 0.02 yuan/kilogram, every plumage meat duck input cost is 0.15 yuan.Thus, every plumage meat duck net output increased value 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 (3)
1. utilize Bacillus licheniformis to produce a method for fodder additives, it 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 be 10
-8time, solid plate substratum carries out coating to it and is separated or line separation, picking part list bacterium colony carries out the mensuration of throughput; Through repeated screening, determine that active strong, that viable count the is high bacterial strain of the gemma produced through fermentation replaces original bacterial strain;
(2) slant strains makes
By recipe requirements configuration slant medium, each component weight part of slant medium is: extractum carnis 5 g, peptone 10 g, NaCl 5 g, and agar powder 20-25 g, adds water and be settled to 1000 mL; PH controls at 7.2-7.4;
Each component is fully dissolved slant medium heating, mix up pH, be then sub-packed in some test tubes, sealing test tube; By test tube in 121 DEG C of sterilizing 30 min, take out pendulum inclined-plane, for subsequent use after cooling;
Above-mentioned slant medium is placed in loft drier or incubator to place 24-48h and access bacterial classification prepared by step (1), wherein Bacillus licheniformis inoculum size is 1% of mixture total weight amount in test tube, under 37 DEG C of conditions, cultivate 24h, bevel bacterial classification;
(3) triangular flask liquid spawn makes
By recipe requirements configuration liquid nutrient medium, the each component weight part of liquid nutrient medium is: peptone 2 g, extractum carnis 3 g, yeast extract paste 2 g, manganous sulfate 0.2 g, potassium primary phosphate 0.4 g, dipotassium hydrogen phosphate 0.5 g, sucrose 20 g, add water and be settled to 1000mL; PH controls at 7.0-7.2; Be sub-packed in the triangular flask of several 500mL, often propping up triangular flask loading amount is 220mL; After sealing at 121 DEG C of temperature sterilizing 30min, cooling after for subsequent use;
Aseptically, the slant strains that liquid nutrient medium access step (2) in each triangular flask is obtained, wherein Bacillus licheniformis inoculum size is 10% of mixture total weight amount in triangular flask, carries out shaking table cultivation to triangular flask, temperature controls at 37 DEG C, and rotating speed is 180-200 r/min; Shaking table is obtained triangular flask liquid spawn after cultivating 24h, for subsequent use;
(4) Cans solid fermentation
By recipe requirements configuration solid fermentation substratum, each component weight part of solid fermentation substratum is: brown sugar 0.5%, lime 0.5%, sterilized water 45%, and all the other are wheat bran;
Be distributed into after being mixed evenly by solid fermentation substratum in some Cans, bottleneck newspaper covers, tight by linear system; To above-mentioned Cans sterilizing 60 min at the temperature of 121 DEG C;
After cooling, the liquid spawn that access step (3) is produced, wherein Bacillus licheniformis inoculum size is 5% of mixture total weight amount; 60 h are cultivated between the cultivation of putting into 37 DEG C, when spore forming rate reaches more than 90% fermentation ends, discharging;
Fermented in fermentation porcelain dish further by obtained material, leavening temperature controls at 37 DEG C, and incubation time is 60 h.
2. a kind of method utilizing Bacillus licheniformis to produce fodder additives as claimed in claim 1, is characterized in that: the sealing in described step (2) refers to and cover tampon on test tube, and tampon and test tube notch portion are wrapped newspaper, tight by linear system.
3. a kind of method utilizing Bacillus licheniformis to produce fodder additives as claimed in claim 1, is characterized in that: the loading amount of often propping up slant medium in test tube in step (2) is the 1/3-1/2 of test tube total volume.
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| CN103911324B (en) * | 2014-03-28 | 2016-05-25 | 内蒙古和美科盛生物技术有限公司 | Contain probiotics preparation of Lactobacillus plantarum and bacillus licheniformis and preparation method thereof |
| CN103952345B (en) * | 2014-04-17 | 2017-01-11 | 青岛九和宜生生物科技有限公司 | Probiotic preparation containing Lactobacillus casei and Bacillus licheniformis and preparation method thereof |
| CN103937715B (en) * | 2014-04-17 | 2017-03-29 | 青岛九和宜生生物科技有限公司 | Tiny ecosystem probiotics preparation containing lactobacillus casei and preparation method thereof |
| CN104543401B (en) * | 2015-01-20 | 2018-04-20 | 河南科技大学 | A kind of preparation method for the state manganese feed addictive that ferments |
| CN105661012B (en) * | 2016-03-09 | 2019-10-29 | 广东海洋大学 | A kind of hot fermentation method of Chinese herbal medicine |
| CN110157636A (en) * | 2019-04-01 | 2019-08-23 | 成都市农林科学院 | A kind of bacillus licheniformis, screening technique, the feed using and containing the bacillus licheniformis |
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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 |