CN105265790B - The cassava alcohol dreg fodder and the preparation method and application thereof of multiple-microorganism fermentation - Google Patents
The cassava alcohol dreg fodder and the preparation method and application thereof of multiple-microorganism fermentation Download PDFInfo
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- CN105265790B CN105265790B CN201510785731.6A CN201510785731A CN105265790B CN 105265790 B CN105265790 B CN 105265790B CN 201510785731 A CN201510785731 A CN 201510785731A CN 105265790 B CN105265790 B CN 105265790B
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Abstract
The invention discloses a kind of liquid bacterium solutions for fermentation cassava alcohol dreg fodder, it is being composed by one or both of Bacillus and lactic acid bacteria；Wherein Bacillus and lactobacillus inoculum ratio are 3:2（v/v）, Bacillus B1 is 1 with Bacillus B2 inoculative proportions:2（v/v）, lactic acid bacteria R1 and Bacillus R2 inoculative proportions are 1:2‑3（v/v）.The present invention further discloses the liquid bacterium solutions for fermentation cassava alcohol dreg fodder to prepare improvement feed flavor, improves the application in terms of palatability.Experimental result is shown：The prebiotic bacterium number of cassava alcohol dreg fodder of liquid bacterium solution fermentation using the present invention significantly improves.Cassava alcohol dreg fodder is obviously increased through everfermentation, lactic acid content.Cassava alcohol dreg fodder through everfermentation has apparent sour-sweet smell.
Description
Technical field
The invention belongs to technical field of microbial fermentation, are related to the cassava alcohol dreg fodder and its system of multiple-microorganism fermentation
Preparation Method and application.
Background technology
Cassava alcohol slag be using large agricultural and sideline product caused by cassava production alcohol, mainly by the skin outside cassava,
Internal parenchymal tissue and cyanogen glycoside material composition, containing a large amount of crude fibre, but crude protein content is relatively low, degree of lignification
Height, palatability are very poor.Crude fibre has stronger stability, and animal digestion utilization rate is low, poor directly as feed efficiency.Wood
Potato application is strong, and with the increase of the market demand, cassava alcohol slag is also being continuously increased, and the storage time the long more can be made to environment
At pollution, being exposed in nature can go mouldy, and the waste water and noxious material of generation can not only cause soil, compared with havoc, to make
At the health that can also further threaten humans and animals while economic loss.These cassava alcohol slags how are handled to become to restrict
The bottleneck of cassava processing enterprise development.
Cassava alcohol slag crude fiber content is high, and carbohydrate content is high, but protein content is low, it is considered that is one
It kind derives from a wealth of sources, is cheap, bum energy feed.Using biologic treating technique, in bacillus, the phases such as lactic acid bacteria
Under the comprehensive function for closing microorganism, the biological chemistry action of a series of complexity is carried out, the physicochemical properties of manioc waste are changed
Cassava alcohol slag is set to become a kind of potential resource, it will to have vast potential for future development.In recent years, many scholars research shows that：
Utilize the nutritional ingredient in cassava alcohol slag that can improve this kind of feed by the characteristic of microorganism itself or the enzyme of generation
Nutritive value, digestibility and palatability.Using microbial fermentation cassava alcohol slag, can not only accomplish largely to consume cassava alcohol
In addition slag is also a kind of inexpensive method for solving domestic market ruminant feed and lacking, has great development potentiality.
But fermentation cassava dreg fodder mostly uses greatly the microbiological treatment of single culture, such as ensiling at present, nutritive value improves not
More, practical application is also unsatisfactory.And take two or more mixed bacteria handle manioc waste research it is few in number and by
Bad in strain combination, strain survival rate is not high, and production technology is complicated under actual production conditions, of high cost can also limit cassava
The large-scale production of residue fermented feed.Therefore, continue to inquire into efficient exploitation, low cost, can be achieved that manioc waste life is factory produced
The best mixed bacteria and its best technological condition for fermentation of object feed are of great significance.
Invention content
The invention discloses a kind of liquid bacterium solutions for fermentative degradation cassava alcohol dreg fodder, it is characterised in that it be by
One or both of Bacillus and lactic acid bacteria are composed；Wherein Bacillus and lactobacillus inoculum ratio are 3:2（v/v）,
Bacillus B1 is 1 with Bacillus B2 inoculative proportions:2（v/v）, lactic acid bacteria R1 and Bacillus R2 inoculative proportions are 1:2-3（v/v）；
The Bacillus B1 refers to bacillus subtilis 1.1413（CGMCC）, B2 refers to bacillus licheniformis
1.265（CGMCC）.Lactic acid bacteria R1 refers to lactobacillus plantarum 1.557（CGMCC）, B2 refers to bacillus coagulans 1.3220
（CGMCC）.
The present invention further discloses the preparation methods using liquid bacterium solution fermentation cassava alcohol dreg fodder, it is characterised in that
It is carried out by following step：
（1）The raw material proportioning of fermented feed is：Cassava alcohol slag 40% ~ 80%（w/w）, wet method sugar residue 10% ~ 30%（w/w）、
Maize peel 8% ~ 18%（w/w）；
（2）In step（1）Middle addition liquid bacterium solution 3% ~ 8%（w/w）, it is 1% ~ 3% that calcium carbonate additive amount, which is then added,（w/
w）, pH to 5.0 ~ 6.0,28 DEG C ~ 32 DEG C of fermentation temperature are adjusted, fermentation time is 4 days ~ 10 days, obtains the fermentation of cassava alcohol slag and raises
Material；The fermented feed being prepared is prepared directly as feed, or directly as feedstuff as feed.
The liquid bacterium solution refers to：Bacillus and lactic acid bacteria mixed proportion are 3:2（v/v）, Bacillus B1's and B2
Ratio is 1:2（v/v）, the ratio of lactic acid bacteria R1 and R2 are 1:2-3（v/v）.
Cassava alcohol slag of the present invention be using cassava processing alcohol remained by residue, mainly by cassava outside
The skin of portion's brown, internal parenchymal tissue and cyanogen glycoside material composition；The wet method sugar residue is corn extraction grape sugar and starch
Byproduct afterwards；The maize peel be will corn particle impregnate, it is broken after, the corn epidermis separated by washing, water squeezing,
Baking operation is process.
The present invention is further disclosed is preparing improvement feeding for the liquid bacterium solution of fermentative degradation cassava alcohol dreg fodder
Expect flavor, improves the application in terms of palatability.The experimental results showed that：
（1）The prebiotic bacterium number of cassava alcohol dreg fodder fermented using liquid bacterium solution of the present invention is significantly improved.
（2）Cassava alcohol dreg fodder is obviously increased through everfermentation, lactic acid content.
（3）Cassava alcohol dreg fodder through everfermentation has apparent sour-sweet smell.
To achieve the above object, the present invention has mainly done following work：
（1）Determine that fermentation raw material matches
Utmostly to consume using cassava alcohol slag, cassava alcohol slag adding proportion is 40% ~ 80%（w/w）.Due to water
Point excessively high, material composition is unstable and excessive containing miscellaneous bacteria, needs to add some ingredients and stablize relatively and conducive to microorganism attachment
Matrix, it is very low additionally, due to cassava alcohol slag pH, it is unfavorable for thalli growth, needs to add buffer calcium carbonate to adjust pH,
Additive amount is 1% ~ 3%（w/w）.The raw material proportioning of fermented feed is cassava alcohol slag 40% ~ 80% after improvement（w/w）, wet method sugar residue
10%~30%（w/w）, maize peel 8% ~ 18%（w/w）.
（2）The screening of fermenting microbe
Totally 15 plants of the bacterial strain that this research is used, strain category can be divided into 9 plants of brood cell bacterium, be bacillus licheniformis respectively
1.265（CGMCC）, bacillus licheniformis 1.813（CGMCC）, bacillus amyloliquefaciens 1.769（CGMCC）, solution starch gemma bar
Bacterium 1.398（CGMCC）, bacillus subtilis 1.1413（CGMCC）, bacillus subtilis 1.173（CGMCC）, bacillus subtilis
Bacterium 1.836（CGMCC）, bacillus subtilis 1.215（CGMCC）, feed series bacillus 1.3772（CGMCC）.Lactic acid bacteria 6
Strain, is lactobacillus plantarum 1.557 respectively（CGMCC）, lactobacillus plantarum 1.2158（CGMCC）, lactobacillus plantarum 1.555
（CGMCC）, lactobacillus acidophilus 1.1854（CGMCC）, Lactobacillus brevis 1.214（CGMCC）, bacillus coagulans 1.3220
（CGMCC）.
The screening of 2.1 gemma bacteria strains
By producing amylase experiment and biocidal property experiment filters out from 9 plants of brood cell bacteriums high yield amylase and biocidal property is strong
Bacterial strain.
In the conical flask for respectively taking the 250ml that 9 plants of one rings of Bacillus strain are connected to liquid amount 50ml, 37 DEG C of culture 12h, liquid
Body culture medium is：Beef extract 0.5%, peptone 1%, NaCl 0.5%, pH7.2 ~ 7.4.
Activated brood cell bacterium is put respectively and is connected on starch culture-medium tablet, 37 DEG C of constant temperature biochemical cultures are then placed in
It is cultivated in case, whether bacterial strain around have transparent circle generation, the bacterial strain that picking generates transparent circle activates again, and dilution spread arrives if observing
On tablet, it is allowed to generate single bacterium colony, observation generates the size of transparent circle, and the big bacterial strain of picking transparent circle activates preservation
Activated Escherichia coli and staphylococcus aureus are diluted to be mixed into after a certain concentration and have been cooled to 50 DEG C of left sides
In right solid medium, it is down flat plate after mixing.Then larger transparent circle is generated on starch culture-medium by activated
Bacterial strain point be connected on tablet, be placed on 37 DEG C of constant temperature incubations, see whether generate inhibition zone, picking generate inhibition zone bacterial strain again
Secondary activation on dilution spread to tablet, is allowed to generate single bacterium colony, the size of inhibition zone produced by observing, picking inhibition zone it is big 5
Strain bacterial strain purification storage, is bacillus licheniformis 1.265 respectively（CGMCC）, bacillus amyloliquefaciens 1.769（CGMCC）, withered grass
Bacillus 1.1413（CGMCC）, bacillus subtilis 1.836（CGMCC）, feed series bacillus 1.3772（CGMCC）.
The screening of 2.2 lactic acid bacteria strains
It is screened from 6 strains of lactic acid bacteria by lactic acid bacteria lactic acid producing performance test and produces the high lactic acid bacteria of acid.
Activated lactic acid bacterial liquid is inoculated according to 5% ratio in feed respectively, feed is then stirred and is allowed to abundant
Mixing is fitted into after fermentation bag is placed in 37 DEG C of environment the 3d that ferments and is detected.5g fermented feeds are weighed when detection is put into triangular flask
In, 45ml distilled water is added and vibrates 20min, is then centrifuged for taking 10ml supernatants, be surveyed with bio-sensing analyzer after 10 times of dilution
Its fixed lactic acid content chooses 4 plants of high bacterial strain purification storages of lactic acid content, is lactobacillus plantarum 1.557 respectively（CGMCC）, plant
Object lactobacillus 1.2158（CGMCC）, Lactobacillus brevis 1.214（CGMCC）, bacillus coagulans 1.3220（CGMCC）.
The screening of 2.3 suitable solid state fermentation bacterial strains
Feed fermentation lactic acid bacteria is determined first by single bacterium fermentation test, and lactic acid bacteria after activation is divided according to 2% inoculum concentration
Lactic acid bacteria number and lactic acid content Jie Ru not be detected after 3 days in fermentation material, finally select that bacterium number is more, the high lactic acid bacteria of lactic acid content
R1（Lactobacillus plantarum 1.557, CGMCC）With lactic acid bacteria R2（Bacillus coagulans 1.3220, CGMCC）；Then by two plants of lactic acid
Bacterium detects gemma bacterium number, lactic acid content finally determines bud after fermentation respectively and in wherein one plant of Bacillus access fermentation material
Spore bacterium is Bacillus B1（Bacillus licheniformis 1.265, CGMCC）, Bacillus B2（Bacillus subtilis 1.1413, CGMCC）.
（3）The optimization of fermentation condition
The optimization of 3.1 fermented feed Bacillus and lactic acid bacteria mixed proportion
Filter out 2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration
It is 5%, Bacillus and lactobacillus inoculum ratio are respectively 1:2,2:1,1:1,3:1,3:2,2 plants of Bacillus inoculative proportions are 1:1,2
Strains of lactic acid bacteria inoculative proportion is 1:It ferments in 1,32 DEG C of constant temperature biochemical cultivation case after 4d, detects lactic acid content therein and probiotics
The indexs such as number
Lactic acid content and prebiotic bacterium number after the fermentation of 1 different strain mixed proportion of table
As can be seen from Table 1, when the mixed proportion of Bacillus and lactic acid bacteria is 3:When 2, lactic acid content and prebiotic bacterium number reach
To highest, respectively 1.70% and 1.52 × 109cfu/g.Therefore, the mixed proportion of Bacillus and lactic acid bacteria selection is 3:2.
The optimization of 3.2 2 plants of Bacillus ratios of fermented feed
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 inoculative proportions are respectively 3:1,2:1,1:1,1:2,1:3,2
Strains of lactic acid bacteria inoculative proportion is 1:It ferments in 1,32 DEG C of constant temperature biochemical cultivation case after 4d, detects lactic acid content therein and probiotics
The indexs such as number.
Lactic acid content and prebiotic bacterium number after 2 two kinds of Bacillus different mixing proportion fermentations of table
As can be seen from Table 2, the mixed proportion of Bacillus B1 and B2 is 1:2 and 1:1 and 1:3 experimental groups are compared, probiotics
Number difference is simultaneously little, but lactic acid content is substantially higher.Therefore, finally determine that the mixed proportion of 2 plants of Bacillus B1 and B2 are 1:2.
The optimization of 3.3 fermented feed, 2 strains of lactic acid bacteria ratio
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 inoculative proportions are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios point
It Wei 3:1,2:1,1:1,1:2,1:It ferments in 3,32 DEG C of constant temperature biochemical cultivation cases after 4d, detects lactic acid content therein and benefit
The indexs such as aerobic plate count.
Lactic acid content and prebiotic bacterium number after 3 two kinds of lactic acid bacteria different mixing proportion fermentations of table
As can be seen from Table 3, the mixed proportion of two kinds of lactic acid bacterias R1 and R2 are 1:3 experimental group and 1:2 compare, it is prebiotic
There is no too big difference for bacterium number, but lactic acid content wants high compared with the latter.Therefore, the mixed proportion of 2 strains of lactic acid bacteria R1 and R2 determines
It is 1:3.
The optimization of 3.4 fermented feed inoculum concentrations
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, Bacillus and lactic acid bacteria
Inoculative proportion is 3:2,2 plants of Bacillus B1 and B2 ratios are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios are 1:3, inoculum concentration is respectively
1%, 3%, 5%, 7%, 9%, in 32 DEG C of constant temperature biochemical cultivation cases after fermentation 4d, detect the fingers such as lactic acid content therein and prebiotic bacterium number
Mark.
Lactic acid content and prebiotic bacterium number after the fermentation of 4 different vaccination amount of table
As can be seen from Table 4, with the increase of inoculum concentration, lactic acid content is also increasing, and maintains to stablize when reaching 2%, benefit
Aerobic plate count highest when inoculum concentration is 5% is 1.29 × 109Cfu/g is then continuously decreased, and illustrating that lactic acid content is excessively high can inhibit
The growth of thalline.Therefore, ferment effect is best when inoculum concentration is 5%.
The optimization of 3.5 fermented feed temperature
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 ratios are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios are 1:3,
It is respectively placed in 24 DEG C, 28 DEG C, 32 DEG C, 36 DEG C, ferment in 40 DEG C of constant temperature biochemical cultivation cases 4d, detects lactic acid content therein
With the indexs such as prebiotic bacterium number.
Lactic acid content and prebiotic bacterium number after 5 different fermentations temperature fermentation of table
As can be seen from Table 5, when fermentation temperature is 32 DEG C and 36 DEG C, lactic acid content and prebiotic bacterium number are all relatively high, and
Difference is simultaneously little, it is contemplated that therefore energy consumption problem selects fermentation temperature for 32 DEG C.
（4）Optimization of orthogonal test fermentation condition
The bacterial strain that will be filtered out, 2 plants of Bacillus and 2 strains of lactic acid bacteria activated for 2 generations, were carried out with the condition of single factor test optimization orthogonal
Experiment, using L9(33) three level of type three way crossover experiment to the inoculative proportion of Bacillus and lactic acid bacteria, inoculum concentration, fermentation temperature
Degree is analyzed.It ferments after 4d, measures its lactic acid content, moisture, reduced sugar, coliform number, gemma bacterium number and lactic acid bacteria number, with
It is highest as final feed fermentation condition to choose lactic acid content as index for lactic acid content.
6 orthogonal experiment influence factor of table and horizontal list
7 orthogonal experiment range analysis table of table
8 orthogonal experiment analysis of variance table of table
R values are intuitively analyzed from Orthogonal experiment results it is found that the factor for influencing test result is followed successively by C>A>B, i.e., to galactopoiesis
Sour influence factor is followed successively by fermentation temperature, inoculative proportion, inoculum concentration.
The lactic acid that can be seen that A2B2C3 fermentation generations from K values is most, i.e., inoculative proportion is 3:2, inoculum concentration 5%, hair
36 DEG C of ferment temperature.But in the actual production process, it is contemplated that the problem of energy consumption cost, fermentation temperature is determined as 32 DEG C, is optimal
Option.
Pass through the variance analysis of orthogonal experiment, it can be seen that：Factor C（Fermentation temperature）With factor A（Inoculative proportion）To reality
Test the influence significant difference of result, and factor B（Inoculum concentration）Influence difference to experimental result is not notable.
Bacillus and lactic acid bacteria mixed proportion will be used for 3:2, the ratio of Bacillus B1 and B2 are 1:2, lactic acid bacteria R1 and
The ratio of R2 is 1:3 strain is formulated bacterium solution, and liquid spawn additive amount is 5%, and 32 DEG C of fermentation temperature, fermentation time is to obtain for 4 days
Cassava alcohol residue fermented feed, using SBA-40C type bio-sensing analyzers, high performance liquid chromatography（Agilent 1200）Equal instrument
Device analyzes the changes of contents of every ingredient in product after detection is fermented, and the results are shown in Table 9：
As can be seen from Table 9, after everfermentation, lactic acid content and prebiotic bacterium number improve a lot, and pH value also has bright
Aobvious decline is not only able to maintain the stabilization of fermented feed ingredient, and can effectively inhibit the growth of harmful miscellaneous bacteria.
Specific implementation mode
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, various changes or change to material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.
The wherein described cassava alcohol slag, wet method sugar residue, maize peel are industrial large by-product, cassava alcohol slag
It is using the residue remained by cassava processing alcohol, wet method sugar residue is the byproduct after corn extraction grape sugar and starch；
Maize peel be will corn particle impregnate, it is broken after, the corn epidermis separated process by washing, water squeezing, baking operation and
At.
Embodiment 1
Determine that fermentation raw material matches
Cassava alcohol slag additive amount is respectively 50%, 60%, 70%, and the additive amount of wet method sugar residue is respectively 10%, 20%, 30%,
The additive amount of maize peel is respectively 8%, 18%, 28%, and calcium carbonate additive amount is 2%, is combined experiment, finally determines fermented feed
Raw material proportioning be cassava alcohol slag 60%（w/w）, wet method sugar residue 20%（w/w）, maize peel 18%（w/w）, add buffer carbonic acid
Calcium adjusts pH, additive amount 2%（w/w）.
Embodiment 2
The screening of fermenting microbe
The screening of 2.1 gemma bacteria strains
By producing amylase experiment and biocidal property experiment filters out from 9 plants of brood cell bacteriums high yield amylase and biocidal property is strong
5 plants of bacterial strain is bacillus licheniformis 1.265 respectively（CGMCC）, bacillus amyloliquefaciens 1.769（CGMCC）, bacillus subtilis
Bacterium 1.1413（CGMCC）, bacillus subtilis 1.836（CGMCC）, feed series bacillus 1.3772（CGMCC）.
The screening of 2.2 lactic acid bacteria strains
It is screened from 6 strains of lactic acid bacteria by lactic acid bacteria lactic acid producing performance test and produces 4 plants high of lactic acid bacteria of acid, be to plant respectively
Object lactobacillus 1.557（CGMCC）, lactobacillus plantarum 1.2158（CGMCC）, Lactobacillus brevis 1.214（CGMCC）, condensation gemma bar
Bacterium 1.3220（CGMCC）.
2.3 suitable solid state fermentation bacterial strain screenings
The 4 plants of high lactic acid bacterias of production acid filtered out are screened again by single bacterium fermentation test first, determine feed fermentation
Lactic acid bacteria is lactic acid bacteria R1（Lactobacillus plantarum 1.557, CGMCC）With lactic acid bacteria R2（Bacillus coagulans 1.3220, CGMCC）.
Then the wherein one plant access by two strains of lactic acid bacteria respectively with the 5 plants of strong Bacillus of high yield amylase and biocidal property filtered out is sent out
In ferment material, gemma bacterium number is detected after fermentation, and lactic acid content is final to determine that Bacillus is Bacillus B1（Bacillus licheniformis
1.265 CGMCC）, Bacillus B2（Bacillus subtilis 1.1413, CGMCC）.
Embodiment 3
The optimization of fermentation condition
The optimization of 3.1 fermented feed Bacillus and lactic acid bacteria mixed proportion
Filter out 2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration
It is 5%, Bacillus and lactobacillus inoculum ratio are respectively 3:1,3:2,1:1,2 plant of Bacillus inoculative proportion is 1:1,2 strains of lactic acid bacteria
Inoculative proportion is 1:It ferments in 1,32 DEG C of constant temperature biochemical cultivation case after 4d, detects the fingers such as lactic acid content therein and prebiotic bacterium number
Mark.Finally determine that the mixed proportion selection of Bacillus and lactic acid bacteria is 3 according to result:2.
The optimization of 3.2 2 plants of Bacillus ratios of fermented feed
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 inoculative proportions are respectively 1:1,1:2,1:3,2 strains of lactic acid bacteria
Inoculative proportion is 1:It ferments in 1,32 DEG C of constant temperature biochemical cultivation case after 4d, detects the fingers such as lactic acid content therein and prebiotic bacterium number
Mark.Finally determine that the mixed proportion of 2 plants of Bacillus B1 and B2 are 1 according to result:2.
The optimization of 3.3 fermented feed, 2 strains of lactic acid bacteria ratio
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 inoculative proportions are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios point
It Wei 1:2,1:3,1:It ferments in 4,32 DEG C of constant temperature biochemical cultivation cases after 4d, detects lactic acid content therein and prebiotic bacterium number etc.
Index.Finally determine that the mixed proportion of 2 strains of lactic acid bacteria R1 and R2 is 1 according to result:3.
The optimization of 3.4 fermented feed inoculum concentrations
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, Bacillus and lactic acid bacteria
Inoculative proportion is 3:2,2 plants of Bacillus B1 and B2 ratios are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios are 1:3, inoculum concentration is respectively
3%, 5%, 7%, in 32 DEG C of constant temperature biochemical cultivation cases after fermentation 4d, detect the indexs such as lactic acid content therein and prebiotic bacterium number.According to
As a result finally determine that ferment effect is best when inoculum concentration is 5%.
The optimization of 3.5 fermented feed temperature
2 plants of Bacillus and 2 strains of lactic acid bacteria were activated into for 2 generations, are accessed in the fermentation material being pre-mixed, inoculum concentration 5%, bud
Spore bacterium and lactobacillus inoculum ratio are 3:2,2 plants of Bacillus B1 and B2 ratios are 1:2,2 strains of lactic acid bacteria R1 and R2 ratios are 1:3,
It is respectively placed in 28 DEG C, 32 DEG C, ferment in 36 DEG C of constant temperature biochemical cultivation cases 4d, detects lactic acid content therein and prebiotic bacterium number
Etc. indexs.Finally determine that fermentation temperature is 32 DEG C according to result
Embodiment 4
Optimization of orthogonal test fermentation condition
The bacterial strain that will be filtered out, 2 plants of Bacillus and 2 strains of lactic acid bacteria activated for 2 generations, were carried out with the condition of single factor test optimization orthogonal
Experiment, using L9(33) three level of type three way crossover experiment to the inoculative proportion of Bacillus and lactic acid bacteria, inoculum concentration, fermentation temperature
Degree is analyzed.It ferments after 4d, measures its lactic acid content, moisture, reduced sugar, coliform number, gemma bacterium number and lactic acid bacteria number, with
It is highest as final feed fermentation condition to choose lactic acid content as index for lactic acid content.Final result, which is shown, uses gemma
Bacterium and lactic acid bacteria mixed proportion are 3:2, the ratio of Bacillus B1 and B2 are 1:2, the ratio of lactic acid bacteria R1 and R2 are 1:3 strain
It is formulated bacterium solution, liquid spawn additive amount is 5%, and the cassava alcohol residue fermented feed end lactic acid content that 32 DEG C of fermentation temperature obtains is most
Height, quality are best.
Contrast test
Claims (4)
1. a kind of liquid bacterium solution for fermentation cassava alcohol dreg fodder, it is characterised in that it is combined by Bacillus and lactic acid bacteria
It forms；Wherein Bacillus and lactic acid bacteria mixed proportion are 3:2（v/v）, Bacillus B1 is 1 with Bacillus B2 mixed proportions:2（v/
v）, lactic acid bacteria R1 and lactic acid bacteria R2 mixed proportions are 1: 3（v/v）；
The Bacillus B1 refers to that bacillus subtilis CGMCC 1.1413, Bacillus B2 refer to bacillus licheniformis
CGMCC 1.265, lactic acid bacteria R1 refer to that lactobacillus plantarum CGMCC 1.557, lactic acid bacteria R2 refer to bacillus coagulans
CGMCC 1.3220。
2. a kind of using the liquid bacterium solution fermentation cassava alcohol dreg fodder for being used for fermentation cassava alcohol dreg fodder described in claim 1
Preparation method, it is characterised in that carried out by following step：
（1）The raw material proportioning of fermented feed is：Cassava alcohol slag 40% ~ 80%（w/w）, wet method sugar residue 10% ~ 30%（w/w）, corn
Skin 8% ~ 18%（w/w）；
（2）In step（1）Middle addition liquid bacterium solution 3% ~ 8%（w/w）, calcium carbonate is then added, additive amount is 1% ~ 3%（w/w）, adjust
PH to 5.0 ~ 6.0,28 DEG C ~ 32 DEG C of fermentation temperature are saved, fermentation time is 4 days ~ 10 days, obtains cassava alcohol residue fermented feed；
The liquid bacterium solution refers to：Wherein Bacillus and lactic acid bacteria mixed proportion are 3:2（v/v）, Bacillus B1 and gemma
Bacterium B2 mixed proportions are 1:2（v/v）, lactic acid bacteria R1 and lactic acid bacteria R2 mixed proportions are 1: 3（v/v）.
3. the preparation method described in claim 2, it is characterised in that：The cassava alcohol slag refers to processing using cassava
Residue remained by alcohol, mainly by the skin of brown outside cassava, internal parenchymal tissue and cyanogen glycoside material composition；It is described wet
Method sugar residue is the byproduct after corn extraction grape sugar and starch；The maize peel is separation after corn particle immersion, being crushed
Corn epidermis out is process by washing, water squeezing, baking operation.
4. the liquid bacterium solution described in claim 1 for fermentation cassava alcohol dreg fodder is preparing improvement feed flavor, improve
Application in terms of palatability.
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| CN107173545A (en) * | 2017-06-23 | 2017-09-19 | 广西中粮生物质能源有限公司 | A kind of method that cassava grain stillage prepares fermented feed |
| CN109043122A (en) * | 2018-09-29 | 2018-12-21 | 天津科技大学 | For the liquid bacterium solution of fermentation cassava dreg fodder and its preparation method of Starch Tapioca residue fermented feed |
| CN112369496B (en) * | 2020-10-20 | 2022-08-16 | 珠海市德海生物科技有限公司 | Modified fermented cassava starch and application thereof in aquatic feed |
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