CN102550818A - Clostridium butyricum as well as colstridium butyricum feed additive and preparation method thereof - Google Patents
Clostridium butyricum as well as colstridium butyricum feed additive and preparation method thereof Download PDFInfo
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- CN102550818A CN102550818A CN2012100018383A CN201210001838A CN102550818A CN 102550818 A CN102550818 A CN 102550818A CN 2012100018383 A CN2012100018383 A CN 2012100018383A CN 201210001838 A CN201210001838 A CN 201210001838A CN 102550818 A CN102550818 A CN 102550818A
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Abstract
The invention discloses a method for preparing a clostridium butyricum feed additive and a clostridium butyricum feed additive prepared by the method. The method for preparing the clostridium butyricum feed additive is characterized by adopting clostridium butyricum (the clostridium butyricum is preserved in the China Center of Industrial Culture Collection and the serial number is CICC 20036). The method comprises the following steps: such materials as tryptone, beef extract, yeast cream, glucose, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulphate, calcium chloride, ferrous sulfate, cysteine hydrochloride and the like are prepared into a dedicated liquid culture medium for seeds; strains are grafted into the culture medium for activating seeds; nitrogen source, carbon source and the like are prepared into a solid anerobic fermentation culture medium; and the seed liquid is grafted into the solid anerobic fermentation culture medium for solid state fermentation and then is dried and sieved. By utilizing the cheap nitrogen and carbon sources to form the medium, the invention provides a novel method for large-scale and industrial production of clostridium butyricum; and the method reduces the cost effectively and increases the benefits of plants.
Description
Technical field
The present invention relates to a kind of clostridium butyricum and clostridium butyricum feed addictive, belong to the microbial fermentation field.
Background technology
Clostridium butyricum has another name called clostridium butyricum, is the born of the same parents bacillus that sprouts in a kind of Gram-positive anaerobism.Bud is embraced huge, generates behind the bud robe or an end expands drum hammer shape, or the middle part expand into spindle shape or shuttle shape.To fowl poultry nonhazardous effect; Biological competition repulsive interaction through in the animal alimentary canal can suppress harmful intestinal tract bacteria, promotes the beneficial bacterium growth; Can reduce ammonia, amine content in the intestines, reduce burden of liver; Produce multivitamin and trace element; Produce multiple enzyme, improve efficiency of feed utilization, promote growth of animal.Remarkable as the probiotics effect, and can use with the antibiotic compatibility.Clostridium butyricum mainly utilizes deep liquid to leave standstill culture technique at present, and the method ratio defective product height but production cost is high is high to the requirement of facility environment, and solid state fermentation cultural method is rare appears in the newspapers for it.
Summary of the invention
It is with high costs to the objective of the invention is to overcome prior art, and the inferior position that working condition is harsh provides a kind of suitability for industrialized production cheap, not harsh to working condition, and has the clostridium butyricum and the clostridium butyricum feed addictive of growth promotion function.
In order to achieve the above object, the invention provides the preparation method of a kind of clostridium butyricum and clostridium butyricum feed addictive, it is characterized in that; (culture presevation is in Chinese industrial microorganism fungus kind preservation administrative center to adopt clostridium butyricum; Be numbered CICC20036, plant name butyricum, generic name Clostridium; History derives from Sichuan Food Fermentative Industry Design Academy (1.265)), concrete steps are following:
The first step: with tryptone 2-2.5g, beef extract 1-1.5g, yeast extract 0.6-1.0g, glucose 0.4-0.5g; Dipotassium hydrogen phosphate 0.2-0.3g, potassium dihydrogen phosphate 0.1-0.3g, magnesium sulfate 0.04-0.06g; Calcium chloride 0.02-0.05g, ferrous sulfate 0.01-0.05g, cysteine hydrochloride 0.05-0.1g; Distilled water 100ml is mixed with the liquid seeds special culture media, and pH value is adjusted to 7.2-7.4; The clostridium butyricum bacterial classification of cold storage is inserted culture medium, and 37-40 ℃ leaves standstill anaerobism cultivation 18-25h, carries out seed activation;
Second step: by percentage to the quality nitrogenous source 20-30%, carbon source 10-15%, calcium carbonate 0.05-0.1%, magnesium sulfate 0.05-0.1%, calcium chloride 0.1-0.2% and remaining water are mixed with the solid anaerobic digestion culture medium;
The 3rd step: carry out solid state fermentation in the solid anaerobic digestion culture medium that second step of seed liquor access that the first step is obtained obtains;
The 4th step: the product that the 3rd step was obtained carries out low temperature drying, pulverizes, sieves, and metering, packing makes finished product.
Preferably, the nitrogenous source in described second step is any one or two or more mixtures in rapeseed dregs, dregs of beans, peanut meal, corn germ cake and the cottonseed dregs of rice.
Further, the nitrogenous source in described second step before as solid-state fermentation culture medium, is sterilized earlier, and is carried out pre-treatment with neutral proteinase; Nitrogenous source and water weight ratio are 1:1.5-2, and neutral proteinase joins in the mixture of nitrogenous source and water with 1000-1500 μ g/g, enzymolysis processing 3-5h.
Preferably, the carbon source in described second step is that any one or two or more mixture and mass percent in wheat bran, rice bran and the corn flour are the mixture of 10% molasses.
Preferably, the mass percent of the inoculum concentration of the seed liquor in described the 3rd step is 5-15%, and cultivation temperature is 36-40 ℃, and leaving standstill the solid anaerobic digestion time is 45-55h.
Preferably, the temperature of low temperature drying in described the 4th step is 38-42 ℃, dries to the mass percent of moisture to be lower than 8%, and the said hole order that sieves is 1.0 eye mesh screens.
The present invention utilizes cheap carbon nitrogen source matrix, and a kind of novel clostridium butyricum large-scale industrial production method is provided, and effectively reduces cost, and increases factory's benefit.
Description of drawings
Fig. 1 is the datagram of solid anaerobic fermentation inoculum concentration of the present invention to the influence of clostridium butyricum viable count and gemma concentration;
Fig. 2 is the datagram of solid anaerobic fermentation time of the present invention to the influence of clostridium butyricum viable count and gemma concentration;
Fig. 3 is the datagram of solid anaerobic fermentation temperature of the present invention to the influence of clostridium butyricum viable count and gemma concentration.
The specific embodiment
Through specific embodiment the present invention is described further below.
Select for use dregs of beans as nitrogenous source; The mixture of 90wt% wheat bran and 10wt% molasses is as carbon source.
1, obtaining liq seed special culture media, tryptone 2.5g wherein, beef extract 1.5g, yeast extract 0.6g; Glucose 0.4g, dipotassium hydrogen phosphate 0.2g, potassium dihydrogen phosphate 0.1g; Magnesium sulfate 0.04g, calcium chloride 0.02g, ferrous sulfate 0.01g; Cysteine hydrochloride 0.05g, distilled water 100ml regulates pH 7.2.
2, the clostridium butyricum bacterial classification with cold storage inserts culture medium, and 37 ℃ leave standstill anaerobism cultivation 18h.
3, earlier with the nitrogenous source sterilization, utilize neutral proteinase to carry out pre-treatment; It is 1:2 that nitrogenous source adds water weight ratio, and neutral proteinase adds with 1000 μ g/g in the mixture of nitrogenous source and water, enzymolysis processing 3h.
4, configuration solid anaerobic digestion culture medium, wherein the proportioning of mass percent does, nitrogenous source 25%, carbon source 15%, calcium carbonate 0.05%, magnesium sulfate 0.05%, calcium chloride 0.1%, adds water to 100%.
5, with carrying out solid state fermentation in the seed liquor access step 2 solid anaerobic digestion culture medium that activation has finished in the step 1.The seed liquor inoculum concentration is 10%, and cultivation temperature is 37 ℃, and leaving standstill the solid anaerobic digestion time is 50h.
6, fermented product is dried by the fire to moisture at 40 ℃ of low temperature be lower than 8wt%, pulverize then, using the sieve aperture order is that 1.0 purpose screen clothes sieve, metering, and packing makes finished product.
Selecting dregs of beans and peanut meal weight ratio for use is that the mixture of 1:1 is as nitrogenous source; 90wt% wheat bran/rice bran weight ratio is that the mixture of mixture and 10wt% molasses of 1:1 is as carbon source.
1, obtaining liq seed special culture media, tryptone 2.0g wherein, beef extract 1.0g, yeast extract 1.0g; Glucose 0.5g, dipotassium hydrogen phosphate 0.2g, potassium dihydrogen phosphate 0.1g; Magnesium sulfate 0.04g, calcium chloride 0.02g, ferrous sulfate 0.02g; Cysteine hydrochloride 0.05g, distilled water 100ml regulates pH 7.4.
2, the cold storage bacterial classification is inserted culture medium, 38 ℃ leave standstill anaerobism cultivation 19h.
3, earlier with the nitrogenous source sterilization, utilize neutral proteinase to carry out pre-treatment; It is 1:2 that nitrogenous source adds water weight ratio, and neutral proteinase adds with 1200 μ g/g in the mixture of nitrogenous source and water, enzymolysis processing 3.5h.
4, configuration solid anaerobic digestion culture medium, wherein the proportioning of mass percent is nitrogenous source 30%, carbon source 15%, calcium carbonate 0.1%, magnesium sulfate 0.1%, calcium chloride 0.1%, adds water to 100%.
5, with carrying out solid state fermentation in the seed liquor access step 2 solid anaerobic digestion culture medium that activation has finished in the step 1.The seed liquor inoculum concentration is 15%, and cultivation temperature is 37 ℃, and leaving standstill the solid anaerobic digestion time is 55h.
6, fermented product is dried by the fire to moisture at 40 ℃ of low temperature be lower than 8wt%, pulverize then, using the sieve aperture order is that 1.0 purpose screen clothes sieve, metering, and packing makes finished product.
Select 60 of the two-way cross boars of growing up about 70 ages in days, about body weight 30kg for use, be divided into 2 groups at random and compare experiment.The control group boar is not added this additive, and test group is that 5% ratio is added in the common daily ration with percentage by weight.This additive and common complete feed fully stir during use, can feed after evenly.These article need be deposited in the hermetically drying place, avoid mixing stacking with agricultural chemicals, strong stimulation article etc.Logging is following:
| The comparison project | Control group | Test group |
| Starting weight (kg) | 30.57±0.26 | 30.52±0.23 |
| Eventually heavy (kg) | 77.33±8.35 | 81.10±8.22 |
| Weightening finish (kg) | 47.07±1.09 | 51.22±2.01 |
| Average daily feed intake (kg) | 2.59 | 3.18 |
| Feedstuff-meat ratio | 2.99 | 2.51 |
| Feeding cost (unit/head) | 250.20 | 232．51 |
Claims (6)
1. the preparation method of a clostridium butyricum feed addictive; It is characterized in that (culture presevation is numbered CICC20036 in Chinese industrial microorganism fungus kind preservation administrative center to adopt clostridium butyricum; Plant name butyricum; Generic name Clostridium, history derives from Sichuan Food Fermentative Industry Design Academy (1.265)), concrete steps are following:
The first step: with tryptone 2-2.5g, beef extract 1-1.5g, yeast extract 0.6-1.0g, glucose 0.4-0.5g; Dipotassium hydrogen phosphate 0.2-0.3g, potassium dihydrogen phosphate 0.1-0.3g, magnesium sulfate 0.04-0.06g; Calcium chloride 0.02-0.05g, ferrous sulfate 0.01-0.05g, cysteine hydrochloride 0.05-0.1g; Distilled water 100ml is mixed with the liquid seeds special culture media, and pH value is adjusted to 7.2-7.4; The clostridium butyricum bacterial classification of cold storage is inserted culture medium, and 37-40 ℃ leaves standstill anaerobism cultivation 18-25h, carries out seed activation;
Second step: by percentage to the quality nitrogenous source 20-30%, carbon source 10-15%, calcium carbonate 0.05-0.1%, magnesium sulfate 0.05-0.1%, calcium chloride 0.1-0.2% and remaining water are mixed with the solid anaerobic digestion culture medium;
The 3rd step: carry out solid state fermentation in the solid anaerobic digestion culture medium that second step of seed liquor access that the first step is obtained obtains;
The 4th step: the product that the 3rd step was obtained carries out low temperature drying, pulverizes, sieves, and metering, packing makes finished product.
2. the preparation method of a kind of clostridium butyricum feed addictive as claimed in claim 1 is characterized in that, the nitrogenous source in described second step is any one or two or more mixtures in rapeseed dregs, dregs of beans, peanut meal, corn germ cake and the cottonseed dregs of rice.
3. the preparation method of a kind of clostridium butyricum feed addictive as claimed in claim 2 is characterized in that, the nitrogenous source in described second step before as solid-state fermentation culture medium, is sterilized earlier, and carried out pre-treatment with neutral proteinase; Nitrogenous source and water weight ratio are 1:1.5-2, and neutral proteinase joins in the mixture of nitrogenous source and water with 1000-1500 μ g/g, enzymolysis processing 3-5h.
4. the preparation method of a kind of clostridium butyricum feed addictive as claimed in claim 1; It is characterized in that the carbon source in described second step is that any one or two or more mixture and the mass percent in wheat bran, rice bran and the corn flour is the mixture of 10% molasses.
5. the preparation method of a kind of clostridium butyricum feed addictive as claimed in claim 1; It is characterized in that; The mass percent of the inoculum concentration of the seed liquor in described the 3rd step is 5-15%, and cultivation temperature is 36-40 ℃, and leaving standstill the solid anaerobic digestion time is 45-55h.
6. the preparation method of a kind of clostridium butyricum feed addictive as claimed in claim 1; It is characterized in that; The temperature of low temperature drying in described the 4th step is 38-42 ℃, dries to the mass percent of moisture to be lower than 8%, and the said hole order that sieves is 1.0 eye mesh screens.
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Cited By (6)
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| CN103283972A (en) * | 2013-03-20 | 2013-09-11 | 广州格拉姆生物科技有限公司 | A solid fermentation method for producing the probiotic agent of live Clostridium butyricum |
| CN103392923A (en) * | 2013-07-23 | 2013-11-20 | 上海创博生态工程有限公司 | Microorganism feed additive for improving reproduction ability of animals and preparation method thereof |
| CN106509367A (en) * | 2016-09-29 | 2017-03-22 | 浙江工业大学 | Schwanniomyces occidentalis source nucleic acid feed additive and application thereof |
| CN108048355A (en) * | 2017-12-26 | 2018-05-18 | 武汉新华扬生物股份有限公司 | A kind of clostridium butyricum tunning and clostridium butyricum process for solid state fermentation |
| CN108478603A (en) * | 2018-04-03 | 2018-09-04 | 潍坊华英生物科技有限公司 | A kind of inactivation clostridium butyricum injection |
| CN110804564A (en) * | 2019-11-18 | 2020-02-18 | 江苏三仪生物工程有限公司 | Preparation of clostridium butyricum powder and detection culture medium thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283972A (en) * | 2013-03-20 | 2013-09-11 | 广州格拉姆生物科技有限公司 | A solid fermentation method for producing the probiotic agent of live Clostridium butyricum |
| CN103392923A (en) * | 2013-07-23 | 2013-11-20 | 上海创博生态工程有限公司 | Microorganism feed additive for improving reproduction ability of animals and preparation method thereof |
| CN106509367A (en) * | 2016-09-29 | 2017-03-22 | 浙江工业大学 | Schwanniomyces occidentalis source nucleic acid feed additive and application thereof |
| CN106509367B (en) * | 2016-09-29 | 2019-11-29 | 浙江工业大学 | One primary yeast source nucleic acid feed additive and its application |
| CN108048355A (en) * | 2017-12-26 | 2018-05-18 | 武汉新华扬生物股份有限公司 | A kind of clostridium butyricum tunning and clostridium butyricum process for solid state fermentation |
| CN108478603A (en) * | 2018-04-03 | 2018-09-04 | 潍坊华英生物科技有限公司 | A kind of inactivation clostridium butyricum injection |
| CN108478603B (en) * | 2018-04-03 | 2020-09-04 | 潍坊华英生物科技有限公司 | Inactivated clostridium butyricum injection |
| CN110804564A (en) * | 2019-11-18 | 2020-02-18 | 江苏三仪生物工程有限公司 | Preparation of clostridium butyricum powder and detection culture medium thereof |
| CN110804564B (en) * | 2019-11-18 | 2020-10-09 | 江苏三仪生物工程有限公司 | Preparation of clostridium butyricum powder and detection culture medium thereof |
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