CN106119171A - The anti-microbial formulation of a kind of crop material feed fermentation and fermentation technology thereof - Google Patents
The anti-microbial formulation of a kind of crop material feed fermentation and fermentation technology thereof Download PDFInfo
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Abstract
The present invention relates to anti-microbial formulation and the fermentation technology thereof of a kind of crop material feed fermentation.Bacillus pumilus QH 1 is cultivated respectively in LB liquid culture, bacillus subtilis QH 1, Bacillus licheniformis QH 1, Lactobacillus plantarum QH 1, culture fluid is mixed and made into crop material feed fermentation microorganism formulation according to viable count than for 1:1:1:1 1:1:1:5 respectively；Adding in the aqueous plant crushed stalk feedstuff containing 0.1% 0.5% white sugars according to the ratio of 1% 10%, stir aerobic fermentation 5 12 hours at 25 DEG C, then pack closing anaerobic ferments 12 72 hours, obtains fermented feed.Crop material is fermented by the anti-microbial formulation utilizing the present invention, live pig feeding experiment carried out by the feedstuff made, the straw palatability as feedstuff can be effectively improved, promote that piglet searches for food and grows, feeding the live pig weightening finish raising 7.42% of fermented feed live pig group more non-fermented feed group, feedstuff-meat ratio reduces by 1.43%.
Description
Technical field:
The present invention relates to anti-microbial formulation and the fermentation technology thereof of a kind of crop material feed fermentation, belong to feed microbe
Technical field.
Background technology:
Agricultural crop straw is the by-product of grain-production, and the whole world is often produced crop material per year and is about 20～3,000,000,000 tons, China
As a large agricultural country, crop material resource is the abundantest, and annual production is about 5-6 hundred million tons.The main component of crop material is
Carbohydrate, additionally, crop material is possibly together with a small amount of crude protein, crude fat and some vitamin and mineral.Institute in straw
The carbohydrate contained is mainly cellulose, hemicellulose and lignin, accounts for more than the 70% of straw dry weight.Crop material rich in
Organic, it is possible to be processed further utilizing as animal feed.But owing to crop material is poor, much to the palatability of animal
Lacking the enzyme of degraded cellulose, hemicellulose and lignin in domestic animal body, the only small amounts of straw of China is directly by as feedstuff
Being used by ruminant, major part straw is incinerated, and this most easily causes the pollution of environment.Crop material is through microorganism
After fermentation, its nutritional labeling can be improved, increase palatability, and the resistance against diseases of feeding animals can be improved.Therefore, select properly
Microbe species, set up be suitable for fermentation technology, fermenting crop material to produce stalk feedstuff is crop material
The effective way of comprehensive utilization.
The Chinese invention patent (CN201410154866.8) that Zhao Shu equality is applied at it disclose a kind of by condensing bud
Probiotics of spore bacillus, Lactobacillus plantarum and lactobacillus casei composition and preparation method thereof, this microbial inoculum can be with bean cake, bran
Skin, Semen Maydis powder are substrate, and the macro-molecular protein in substrate is degraded to oligopeptide, and by the anti-nutrient substance in albumen, antigen
Albumen etc. thoroughly decompose, thus improve protein quality and the feeding value of the fermented products such as bean cake.Yao Guoqiang etc. apply at it
Chinese invention patent (CN201410121486.4) in disclose a kind of by Bacillus coagulans HM-09, Bacillus licheniformis
Bean cake, wheat bran etc. are carried out solid by the microbial fermentation preparation of BL-09, lactobacillus casei HM-09 and Lactobacillus plantarum HM-10 composition
The method of body fermentation.The Chinese invention patent (CN201310635322.9) that Cui Guochen etc. apply at it is introduced in feedstuff and add
Fermentation of bacillus subtilis liquid dried object can improve poultry absorbability mesotrophic to feedstuff.Li Zhichuns etc. are in its invention specially
Disclosing a kind of feed microbe additive being made up of Trichoderma viride, Candida utilis and plant lactobacillus in profit, this adds
Adding agent and banana stem leaves is carried out ensiling, the fermentation time preparing ensilage is short, banana stem leaves crude fiber content after silage fermentation
Reducing by 20%, crude protein improves more than 30%.Come course line etc. at its Chinese invention patent applied for (CN201010213978.8)
In disclose a kind of micro-by Lactobacillus plantarum, Pediococcus pentosaceus, aroma-producing yeast, Issatchenkia orientalis and bacillus cereus five kinds
The microbe additive of biotic component, adds lactic acid and the acetic acid content raising of the compound ensilage of microorganism, and crude protein contains
Amount has significantly increase, and ratio adds 25% before ensiling.Wang Depei etc. are at its Chinese invention patent applied for
(CN201010213978.8) one kind of multiple fermentable concentrated feeds and ferment essence feed microbe mother solution and system are described in
Making method, this microorganism mother solution uses many bacterial strains such as Cellumomonas flavigena, bacillus amyloliquefaciens to coordinate anaerobic fermentation feedstuff
Method, can ferment various crop material powder (Semen Maydis, Semen Tritici aestivi, Oryza sativa L., Sorghum vulgare Pers. and seedling of Semen arachidis hypogaeae, Rhizoma Dioscoreae seedling etc.) and cereals, furfur
The mixing of class, bean cake class produces cereals concentrated feed, bran concentrated feed and bean cake class concentrated feed.Therefore, find one to be suitable for
The anti-microbial formulation of crop material fermentation, sets up the fermentation technology being suitable for, and the comprehensive development and utilization for crop material has weight
Want meaning.
Summary of the invention:
It is an object of the invention to overcome the shortcoming of prior art, it is provided that the microorganism of a kind of crop material feed fermentation is joined
Side and fermentation technology thereof, by fermenting the crop material of pulverizing this anti-microbial formulation and application process, thus it is possible to vary
The animal palatability of feedstuff, the absorbability nutrition content of increase feedstuff, promote that the growth of feeding animals, enhancing feed dynamic
The resistance against diseases of thing.
In order to realize foregoing invention purpose, our first isolation identification 8 kinds from the corn straw feedstuff of natural fermentation
Microorganism.Through morphological observation, analysis of physio biochemical characteristics and 16SrDNA sequence analysis, 8 kinds of microorganisms are identified respectively as short
Bacillus pumilus QH-1 (preserving number CGMCC No.12094), bacillus subtilis QH-1 (preserving number CGMCC No.12095),
Bacillus licheniformis QH-1 (preserving number CGMCC No.12096), Lactobacillus plantarum QH-1 (preserving number CGMCC No.12097),
A Shi bacillus cereus (Bacillus aryabhattai), air bacillus cereus (Bacillus aerius), solution starch spore bar
Bacterium (Bacillus amyloliquefaciens) and vegetable bacillus cereus (Bacillus oleronius)；
Secondly, 8 kinds of microorganisms to being separated carry out fermenting experiment according to single or different ratio to crop material, according to
Free amino acid, soluble reducing sugars and soluble protein content in fermented product and the animal feed intake etc. to fermented feed
Index, filters out suitable crop material feed fermentation anti-microbial formulation, and anti-microbial formulation consists of: Bacillus pumilus QH-1
(preserving number CGMCC No.12094), bacillus subtilis QH-1 (preserving number CGMCC No.12095), Bacillus licheniformis QH-
1 (preserving number CGMCC No.12096) and Lactobacillus plantarum QH-1 (preserving number CGMCC No.12097), four kinds of microorganisms are lived
Bacterium number is than for 1:1:1:1-1:1:1:5；
The entitled China Committee for Culture Collection of Microorganisms of depositary institution of the above four kinds of biological material specimens
Common micro-organisms center, depositary institution address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation date January 22 in 2016
Day, Classification And Nomenclature is Bacillus pumilus Bacillus pumilus (CGMCC No.12094)；Bacillus subtilis
Bacillus subtilis(CGMCC No.12095)；Bacillus licheniformis Bacillus licheniformis (CGMCC
No.12096)；Lactobacillus plantarum Lactobacillus plantarum (CGMCC No.12097).
Utilizing the anti-microbial formulation filtered out, the fermentation process step of crop material is as follows:
One, feed fermentation microorganism formulation is prepared
Picking Bacillus pumilus QH-1 (preserving number CGMCC No.12094), bacillus subtilis QH-1 (preservation respectively
Number CGMCC No.12095), Bacillus licheniformis QH-1 (preserving number CGMCC No.12096), Lactobacillus plantarum QH-1 (preservation
Number CGMCC No.12097) single bacterium colony, it is inoculated in 10ml LB fluid medium, 25 DEG C, shaken cultivation 24 hours, picking is planted
The single bacterium colony of thing lactobacillus QH-1 (preserving number CGMCC No.12097), is inoculated in 10ml containing the LB liquid that weight ratio is 2% glucose
In body culture medium, quiescent culture 24 hours, it is thus achieved that the fermentation liquid of 4 kinds of bacterial isolateses, colony counting method measures viable bacteria in fermentation liquid
Sum；With LB fluid medium, different strain fermentation liquid being adjusted to viable count is 3.0 × 109The seed liquor of/ml；
By Bacillus pumilus QH-1, bacillus subtilis QH-1, Bacillus licheniformis QH-1, Lactobacillus plantarum QH-1 kind
Sub-liquid is mixed into feed fermentation microorganism formulation according to the ratio that volume ratio is 1:1:1:1 or 1:1:1:5；
Two, straw feed fermentation
Crushing straw wet weight used ratio for 30%-50% (w/w), is 0.1%-0.5% white sugar (w/ containing weight ratio
W), microorganism formulation addition is weight ratio 1%-10%, at 25 DEG C, and stirring aerobic fermentation 5-12 hour, then pack airtight
Anaerobic fermentation 12-72 hour, obtains fermented feed.
In gained fermented feed, soluble reducing sugars, free amino acid and soluble protein are to be not added with microorganism respectively
3.15-4.30 times of the matched group of formula, 2.30-3.00 times and 1.73-1.89 times.
Crop material is fermented by the anti-microbial formulation utilizing the present invention, and live pig feeding experiment carried out by the feedstuff made, fermentation
Feedstuff is not sent out as fermented feed, the conduct adding equivalent white sugar and bean cake after adding the bean cake that weight ratio is 6% (w/w) again
Ferment feedstuff, feeds live pig with fermented feed and non-fermented feed respectively, feeds fermented feed live pig group more non-fermented feed group
Live pig weightening finish raising 7.42%, feedstuff-meat ratio reduces by 1.43%, can be effectively improved the straw palatability as feedstuff, promotes that piglet is adopted
Food and growth.
Detailed description of the invention:
Below by specific embodiment, the invention will be further elaborated.
Embodiment 1:
One, the preparation of fermentation seed liquid
Picking Bacillus pumilus QH-1 (preserving number CGMCC No.12094), bacillus subtilis QH-1 (preservation respectively
Number CGMCC No.12095), Bacillus licheniformis QH-1 (preserving number CGMCC No.12096), Lactobacillus plantarum QH-1 (preservation
Number CGMCC No.12097) single bacterium colony, it is inoculated in 10ml LB fluid medium, 25 DEG C, shaken cultivation 24h, picking plant breast
The single bacterium colony of bacillus QH-1 (preserving number CGMCC No.12097), is inoculated in the 10ml LB fluid medium containing 2% glucose, quiet
Putting cultivation 24h, it is thus achieved that the fermentation liquid of 4 kinds of bacterial isolateses, colony counting method measures total viable count in fermentation liquid, and result shows, short
The viable count of the viable count in bacillus pumilus QH-1 fermentation liquid is 3.87 × 109/ ml, in bacillus subtilis QH-1 fermentation liquid
Viable count be 5.26 × 109/ ml, the viable count of Bacillus licheniformis QH-1 fermentation liquid is 4.35 × 109/ ml, Lactobacillus plantarum
The viable count of the viable count in QH-1 fermentation liquid is 6.69 × 109/ ml, adjusts different strain fermentation liquid with LB fluid medium
Becoming viable count is 3.0 × 109The seed liquor of/ml；
Two, the solid fermentation of corn straw is tested by different strains proportioning
By Bacillus pumilus QH-1 (preserving number CGMCC No.12094), bacillus subtilis QH-1 (preserving number CGMCC
No.12095), Bacillus licheniformis QH-1 (preserving number CGMCC No.12096), Lactobacillus plantarum QH-1 (preserving number CGMCC
No.12097) seed liquor is mixed into feed fermentation microorganism formulation according to the ratio that volume ratio is 1:1:1:1 or 1:1:1:5, presses
It is inoculated into according to the inoculum concentration that weight ratio is 2% and with the addition of corn stalk powder (the wet weight ratio that weight ratio is 0.1% white sugar
50%), 25 DEG C of stirring aerobic fermentation 5-12h, then pack closing anaerobic fermentation 12-72h, obtains fermented feed；
With same treatment, but not having the corn stalk powder of seed addition liquid is comparison；
Sample and measure the viable count in fermented feed, DNS colorimetric method for determining content of reducing sugar, indenes three according to colony assay
Ketone colorimetric method for determining free aminoacid content, Coomassie brilliant blue G250 colorimetric method for determining soluble protein content, result shows, 4
The seed liquor of kind of antibacterial according to viable count in the feedstuff that volume ratio is the microorganism formulation fermentation that 1:1:1:1 is made into be 2.35 ×
106/ g, content of reducing sugar is 3.32mg/g, and the content of free amino acid is 3.77mg/g, and the content of soluble protein is
1.67mg/g；The seed liquor of 4 kinds of antibacterials is according to viable count in the feedstuff that volume ratio is the microorganism formulation fermentation that 1:1:1:5 is made into
It is 2.95 × 106/ g, content of reducing sugar is 4.51mg/g, and the content of free amino acid is 4.24mg/g, containing of soluble protein
Amount is 1.82mg/g；And be not added with antibacterial seed liquor and in the corn stalk powder of same treatment content of reducing sugar be
1.05mg/g, the content of free amino acid is 1.41mg/g, and the content of soluble protein is 0.96mg/g.
Embodiment 2, the live pig feeding experiment of interpolation fermented feed
Choosing the piglet that body weight is about 30kg, be randomly divided into 2 groups, often group 10, weighs each group of original body mass, and one group is
Control group fed premixing feedstuff, another group feeds the premixing feedstuff that with the addition of weight ratio 10% fermented feed, feeds 4 for test group
Each parameter is added up after individual month.
Result shows, matched group total augment weight is 1079.6kg, and total feed intake is 3024kg, and feedstuff-meat ratio is 2.80；And test
Group total augment weight is 1159.7kg, and total feed intake is 3200kg, and feedstuff-meat ratio is 2.76.Illustrate feedstuff adds a certain amount of institute of the present invention
Fermented feed prepared by the method for stating can improve the palatability of feedstuff, promotes that piglet searches for food and grows, reduces feedstuff-meat ratio.
Claims (3)
1. the anti-microbial formulation of a crop material feed fermentation, it is characterised in that anti-microbial formulation consists of: short and small spore bar
Bacterium QH-1, bacillus subtilis QH-1, Bacillus licheniformis QH-1 and Lactobacillus plantarum QH-1, four kinds of microorganism viable bacteria number ratios
For 1:1:1:1-1:1:1:5；Wherein, Bacillus pumilus QH-1 preserving number is CGMCC No.12094, bacillus subtilis QH-
1 preserving number is CGMCC No.12095, and Bacillus licheniformis QH-1 preserving number is CGMCC No.12096, Lactobacillus plantarum QH-1
Preserving number is CGMCC No.12097.
2. a crop material feed fermenting process, it is characterised in that operation in accordance with the following steps:, prepare the micro-life of feed fermentation
Thing preparation: picking Bacillus pumilus QH-1, bacillus subtilis QH-1, Bacillus licheniformis QH-1, Lactobacillus plantarum respectively
The mono-bacterium colony of QH-1, is inoculated in 10ml LB fluid medium, 25 DEG C, shaken cultivation 24 hours, and Lactobacillus plantarum QH-1 is mono-for picking
Bacterium colony, is inoculated in the LB fluid medium that 10ml is 2% glucose containing weight ratio, quiescent culture 24 hours, it is thus achieved that 4 kinds of antibacterials
The fermentation liquid of bacterial strain, colony counting method measures total viable count in fermentation liquid；With LB fluid medium, different strain fermentation liquid is adjusted
Being made into viable count is 3.0 × 109The seed liquor of/ml；By Bacillus pumilus QH-1, bacillus subtilis QH-1, lichens spore
Bacillus QH-1, Lactobacillus plantarum QH-1 seed liquor are mixed into feed fermentation according to the ratio that volume ratio is 1:1:1:1 or 1:1:1:5
Microorganism formulation；Two, straw feed fermentation: crushing straw wet weight used ratio for 30%-50%, is 0.1%-containing weight ratio
0.5% white sugar, microorganism formulation addition is weight ratio 1%-10%, at 25 DEG C, stirring aerobic fermentation 5-12 hour, then
Pack closing anaerobic fermentation 12-72h, obtains fermented feed.
A kind of crop material feed fermenting process the most according to claim 2, it is characterised in that solubility in fermented feed
Reducing sugar, free amino acid and soluble protein be 3.15-4.30 times of the matched group being not added with anti-microbial formulation respectively,
2.30-3.00 again and 1.73-1.89 times.
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| CN106819380A (en) * | 2016-12-31 | 2017-06-13 | 新昌县田野泉养殖技术开发有限公司 | Biological protein feed and preparation method thereof, application method |
| CN107502572A (en) * | 2017-09-08 | 2017-12-22 | 中国农业科学院农业环境与可持续发展研究所 | Application of the bacillus licheniformis in straw degradative, the microbial bacterial agent comprising the bacterium and its application |
| CN107603924A (en) * | 2017-11-01 | 2018-01-19 | 天津博菲德科技有限公司 | A kind of complex microorganism preparations and its preparation method and application |
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| CN107502572B (en) * | 2017-09-08 | 2020-08-21 | 中国农业科学院农业环境与可持续发展研究所 | Application of bacillus licheniformis in straw degradation, microbial agent containing bacillus licheniformis and application of bacillus licheniformis |
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| CN109965083A (en) * | 2019-04-17 | 2019-07-05 | 佛山播恩生物科技有限公司 | A kind of depth fermented feed production method containing two kinds of high-content organic acids |
| CN110353087A (en) * | 2019-07-11 | 2019-10-22 | 南京农业大学 | The preparation method and fermented feed of rape stalk fermented feed |
| CN110477190A (en) * | 2019-08-28 | 2019-11-22 | 上海交通大学 | A kind of stalk anaerobic fermentation feed and preparation process and formula forage |
| CN111165653A (en) * | 2020-02-19 | 2020-05-19 | 黑龙江八一农垦大学 | Potato residue and corn straw mixture fermented feed and preparation method and leavening agent thereof |
| CN111165653B (en) * | 2020-02-19 | 2023-09-12 | 黑龙江八一农垦大学 | Potato residue and corn stalk mixture fermented feed, preparation method thereof and starter |
| CN112998130A (en) * | 2021-04-22 | 2021-06-22 | 沈阳工学院 | Pomace and crop straw mixed fermented feed and preparation method thereof |
| CN112998130B (en) * | 2021-04-22 | 2023-08-22 | 沈阳工学院 | Fruit residue and crop straw mixed fermentation feed and preparation method thereof |
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