CN105543131A - Compound bacteria/cottonseed meal fermented feed and preparation method thereof - Google Patents

Compound bacteria/cottonseed meal fermented feed and preparation method thereof Download PDF

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Publication number
CN105543131A
CN105543131A CN201511021332.9A CN201511021332A CN105543131A CN 105543131 A CN105543131 A CN 105543131A CN 201511021332 A CN201511021332 A CN 201511021332A CN 105543131 A CN105543131 A CN 105543131A
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China
Prior art keywords
cotton dregs
fermentation
saccharomyces cerevisiae
bacillus licheniformis
fermented feed
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CN201511021332.9A
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Chinese (zh)
Inventor
雷恒
王勇生
程宗佳
陈婧
香红星
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Feed Corp ltd Of China Oil And Food Import And Export Corporation
Cofco Nutrition and Health Research Institute Co Ltd
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Feed Corp ltd Of China Oil And Food Import And Export Corporation
Cofco Nutrition and Health Research Institute Co Ltd
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Priority to CN201511021332.9A priority Critical patent/CN105543131A/en
Publication of CN105543131A publication Critical patent/CN105543131A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • C12N1/16Yeasts; Culture media therefor
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23YINDEXING SCHEME RELATING TO LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23Y2220/00Lactobacillus
    • A23Y2220/67Plantarum
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87Re-use of by-products of food processing for fodder production

Abstract

The invention relates to the technical field of feed science, particularly a compound bacteria/cottonseed meal fermented feed and a preparation method thereof. The invention provides a compound bacteria/cottonseed meal fermented feed and a preparation method thereof. The compound bacteria comprise Lactobacillus plantarum, Bacillus subtilis, Bacillus licheniformis and Saccharomyces cerevisiae. The research detects that when the Lactobacillus plantarum, Bacillus subtilis, Bacillus licheniformis and Saccharomyces cerevisiae are compounded, the fermentation action of the compound bacteria can enhance the contents of cottonseed meal crude proteins, small molecule peptides and lactic acid; and after the fermentation, the crude protein content is at least 49.8%, the small peptide content accounts for at least 9.0% of the crude proteins, and the lactic acid content (based on total acids) is at least 2.2%. Thus, the compound bacteria have obvious better fermentation effects on the cottonseed meal than each single bacterium.

Description

Composite bacteria, cotton dregs fermented feed and preparation method thereof
Technical field
The present invention relates to forage science technical field, particularly composite bacteria, cotton dregs fermented feed and preparation method thereof.
Background technology
Along with the fast development of livestock industry, protein feed resource shortage is more and more serious, how rational exploitation and use protein feed resource, plays vital effect by the development of China's livestock industry.Cotton dregs are face cakes that cottonseed draws after squeezing, separate through most of Residual oil of extract technology by the inside again, a kind of micro-red or yellow granular article obtained, cotton dregs are one of main plant protein feed resources of China, it is the main raw material of Fodder making, and the crude protein contained can reach more than 40%.
But cotton dregs contain the antinutritional factor such as gossypol because of it, and digestive utilization ratio is low, is difficult to a large amount of use.Gossypol is mainly present in cotton benevolence pigment gland, is a kind of water insoluble and the tawny being dissolved in organic solvent gathers phenol pigment.In liquefaction process, owing to steaming the heat effects such as stir-fry, squeezing, most of gossypol is combined with protein, amino acid and becomes bound gossoypol, and bound gossoypol is not absorbed by animal in animal digestive tract, therefore toxicity is very little.Another part gossypol is then present in cake, the dregs of rice and oil product in a free form, this part free gossypol is larger to animal toxicity, especially monogastric animal excess ingestion or picked-up the time longer, growth retardation, reproductive performance and production performance can be caused to decline, even cause death.The tolerance of youngling to gossypol is lower.The toxic dose of free gossypol is relevant with protein level, ferrous ion level and calcium ion level in diet.
Solid microbe fermentation technology is applied comparatively extensive in feedstuff industry, its main purpose improves product quality by the mode of solid state fermentation, improve palatability, reduce the antinutritional factor content in raw material, improve the digestibility of feed, become the feeds product of natural nuisance-free height nutrition.Utilize the microorganisms such as milk-acid bacteria to carry out fermentative processing to cotton dregs, protein macromolecule can be degraded to by the metabolism of microorganism the small-molecule substance that little peptide etc. is easy to absorption; Fermenting process can breed a large amount of beneficial microorganisms simultaneously, and produces the organic acids such as a large amount of lactic acid, reduces pH, suppress the growth of harmful bacteria, and the organic acids such as the lactic acid of fermentation generation make feed have sour fragrance, can improve the palatability of cotton dregs, improve the application addition of cotton dregs in feed.
At present, adopt the ferment effect of single lactic acid bacteria culturers fermentation cotton dregs poor, be badly in need of the better fermentation process of a kind of ferment effect.
Summary of the invention
In view of this, the invention provides a kind of composite bacteria, cotton dregs fermented feed and preparation method thereof.The present invention study find by plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae with the use of, utilize the fermentative action of this composite bacteria, can improve cotton dregs crude protein, the content of small-molecular peptides and lactic acid content, composite bacteria is significantly better than each single bacterium to the ferment effect of cotton dregs to the ferment effect of cotton dregs.
In order to realize foregoing invention object, the invention provides following technical scheme:
The invention provides a kind of composite bacteria, comprise plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
Plant lactobacillus is the one of milk-acid bacteria, and optimum growth temperature is 30 ~ 35, anaerobism or amphimicrobian, bacterial classification be straight or curved shaft-like, single, sometimes in pairs or become chain, optimal pH about 6.5, belongs to homofermentative lactic bacteria.This bacterium and the difference of other milk-acid bacterias are that the viable count of this bacterium is higher, and product acid that can be a large amount of, makes the pH value in water stablize and do not raise, and the acid mass-energy degraded heavy metal of its output; Because this bacterium is anaerobic bacterium (facultative aerobic), the distinctive lactobacillin of energy output in reproductive process, lactobacillin is the sanitas of a kind of biotype.In the cultivation middle and later periods, due to ight soil and the increase of residual bait of animal, can sink to the bottom in pond, and rot, grow a lot of germ, generate a large amount of ammonia nitrogens and nitrite, dead phenomenon is serious steathily to make bottom.If life-time service lactobacillus plantarum, just can well suppress bottom ight soil and residual bait rot, also just reduce the increase of ammonia nitrogen and nitrite, substantially reduce the number the consumption of chemical industry degradation element, aquaculture cost is reduced.
Bacillus subtillis is the one of Bacillus.Individual cells 0.7 ~ 0.8 × 2 ~ 3 microns, uniform coloring.Without pod membrane, peritrichous, can move.Gram-positive microorganism, gemma 0.6 ~ 0.9 × 1.0 ~ 1.5 microns, oval to column, be positioned at thalline central authorities or slightly inclined, after sporulation, thalline does not expand.Bacterium colony surface irregularity is opaque, dirty white or micro-yellow, when growing in liquid medium within, and normal formation wrinkle mould.Aerophil.Available protein, multiple sugar and starch, decompose tryptophane and form indoles.Be widely used in genetics research, to the route of synthesis of the purine nucleotides of this bacterium and its regulation mechanism research clearer.Extensively be distributed in the organism of soil and corruption, easily breed in withered grass leaching juice.
Bacillus licheniformis cell form and arrangement are in shaft-like, Dan Sheng, and adjustable flora imbalance reaches therapeutic purpose, and body can be impelled to produce antibacterial substance, kill pathogenic bacterium.Can produce activity resistent material, and the biology with uniqueness takes oxygen mechanism of action by force, can suppress the growth and breeding of pathogenic bacterium.
Yeast saccharomyces cerevisiae (Saccharomycescerevisiae), also known as bread yeast or budding yeast.Yeast saccharomyces cerevisiae is and human relation's primary yeast the most widely, not only because it is for making the food such as bread and steamed bun and wine brewing traditionally, in modern molecular and cytobiology, be used as eucaryon model animals, its effect is equivalent to the model animals intestinal bacteria of protokaryon.Yeast saccharomyces cerevisiae is biological species the most frequently used in fermentation.The cell of yeast saccharomyces cerevisiae is spherical or avette, diameter 5 ~ 10 μm.The method of its breeding is gemmation.
The present invention study find by plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae with the use of, it is significantly better than each single bacterium to the ferment effect of cotton dregs to the ferment effect of cotton dregs.
As preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is (1 ~ 5): (1 ~ 3): (1 ~ 3): (1 ~ 3).
Preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 3:2:2:2.
In embodiments more provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 1:3:1:3.
In other embodiments provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 5:1:3:1.
Present invention also offers a kind of cotton dregs fermented feed, its preparation method comprises:
Cotton dregs are mixed with water, obtains fermentation material, fermentation material is mixed with composite bacteria, adopt solid state fermentation to ferment, dry, obtain cotton dregs fermented feed;
Composite bacteria comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
As preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is (1 ~ 5): (1 ~ 3): (1 ~ 3): (1 ~ 3).
Preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 3:2:2:2.
In embodiments more provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 1:3:1:3.
In other embodiments provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 5:1:3:1.
As preferably, the mass percentage of water in fermentation material is 45% ~ 65%.
Preferably, the mass percentage of water in fermentation material is 50%.
As preferably, in CFU/g, the consumption of composite bacteria and cotton dregs is (2.5 ~ 10) × 10 8: 1000.
Preferably, in CFU/g, the consumption of composite bacteria and cotton dregs is 5 × 10 8: 1000.
As preferably, the temperature of solid state fermentation is 36 ~ 38 DEG C.
Preferably, the temperature of solid state fermentation is 37 DEG C.
As preferably, the time of solid state fermentation is 1 ~ 6d.
Preferably, the time of solid state fermentation is 4d.
As preferably, ferment and adopt anaerobically fermenting.
As preferably, the temperature of oven dry is 70 ~ 90 DEG C.
Preferably, the temperature of oven dry is 80 DEG C.
Present invention also offers a kind of preparation method of cotton dregs fermented feed, comprise the steps:
Cotton dregs are mixed with water, obtains fermentation material, fermentation material is mixed with composite bacteria, adopt solid state fermentation to ferment, dry, obtain cotton dregs fermented feed;
Composite bacteria comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
As preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is (1 ~ 5): (1 ~ 3): (1 ~ 3): (1 ~ 3).
Preferably, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 3:2:2:2.
In embodiments more provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 1:3:1:3.
In other embodiments provided by the invention, in CFU, the consumption of plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae is 5:1:3:1.
As preferably, the mass percentage of water in fermentation material is 45% ~ 65%.
Preferably, the mass percentage of water in fermentation material is 50%.
As preferably, in CFU/g, the consumption of composite bacteria and cotton dregs is (2.5 ~ 10) × 10 8: 1000.
Preferably, in CFU/g, the consumption of composite bacteria and cotton dregs is 5 × 10 8: 1000.
As preferably, the temperature of solid state fermentation is 36 ~ 38 DEG C.
Preferably, the temperature of solid state fermentation is 37 DEG C.
As preferably, the time of solid state fermentation is 1 ~ 6d.
Preferably, the time of solid state fermentation is 4d.
As preferably, ferment and adopt anaerobically fermenting.
As preferably, the temperature of oven dry is 70 ~ 90 DEG C.
Preferably, the temperature of oven dry is 80 DEG C.
The invention provides a kind of composite bacteria, cotton dregs fermented feed and preparation method thereof.This composite bacteria comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.Beneficial effect of the present invention is:
The present invention study find by plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae with the use of, utilize the fermentative action of this composite bacteria, cotton dregs crude protein, the content of small-molecular peptides and lactic acid content can be improved, after fermentation, gross protein value >=49.8%; Little peptide content (accounting for crude protein) >=9.0%; Lactic acid content (in total acid) >=2.2%.Visible, composite bacteria is significantly better than each single bacterium to the ferment effect of cotton dregs to the ferment effect of cotton dregs.
After compound bacteria-fermented cotton dregs, pH value is lower than pH value after single bacterium fermentation cotton dregs, is easy to preserve.
Embodiment
The invention discloses a kind of composite bacteria, cotton dregs fermented feed and preparation method thereof, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
In composite bacteria provided by the invention, cotton dregs fermented feed and preparation method thereof, bacterial classification used all can be buied by market.Four kinds of bacterial strain compositions in the present invention's composite bacteria used: plant lactobacillus (LactobacillusplantarumZF14), subtilis (BacillussubtilisZF21), Bacillus licheniformis (BacilluslicheniformisZF23), yeast saccharomyces cerevisiae (SaccharomycescerevisiaeZF31), all sell company limited purchased from Dingzhou posivtive spike feed.
Below in conjunction with embodiment, set forth the present invention further:
The impact that embodiment 1 moisture ferments on cotton dregs
1, materials and methods
1.1, bacterial classification
Test composite bacteria used to be made up of plant lactobacillus, subtilis, Bacillus licheniformis, yeast saccharomyces cerevisiae, in CFU, the ratio of above-mentioned four kinds of bacterium is 3:2:2:2.
1.2, cotton dregs raw material sources
Cotton dregs, purchased from China Oil and Food Import and Export Corporation (Changji) cereal and oil industry company limited, are that the face cake of cottonseed after squeezing is separated through most of Residual oil of extract technology by the inside again, a kind of by product obtained.
1.3, test design
Take cotton dregs 200g, add moisture in 45%, 50%, 55%, 60%, 65% ratio of fermentation material total amount, be designated as T respectively 1, T 2, T 3, T 4, T 5.Add composite bacteria 1 × 10 respectively 8cFU, leavening temperature controls at 37 DEG C, fermentation time 4d, 80 DEG C of oven dry after fermentation ends, crosses 40 mesh sieves after crushed, and measures the indexs such as crude protein, little peptide, lactic acid.
1.4, chemical analysis
A, gross protein value detect: perform by GB/T5009.5-2003 regulation.
B, little peptide content detect and adopt trichoroacetic acid(TCA) extraction method: accurately take fermentation cotton dregs sample 1.5g (being accurate to 0.0001g) in 100mL beaker, add 15% solution of trichloroacetic acid 25mL, vibration 5min, leave standstill 5min, solution is quantitatively shifted, at 3,000 rpm centrifugal 10min, get whole supernatant liquid filtering, accurately pipette 10mL filtrate and be placed in alimentary canal, digest by GB/T5009.5-2003 method and measure protein content.Little peptide calculation formula is as follows:
Little peptide (accounting for crude protein, %)=(V-V 0) × C × 6.25 × 0.014 × 2.5 × 100/ (M × CP)
In formula: V-sample consumes the volume of HCl, mL;
V 0-blank the volume consuming HCl, mL;
The volumetric molar concentration of C-hydrochloric acid, mol/L;
M-sample quality, g;
6.25 × 0.014-albumen gain factor;
The gross protein value of CP-fermentation cotton dregs sample.
C, lactic acid content (in total acid) adopt NaOH titration measuring: take 0.5g and ferment cotton dregs sample (being accurate to 0.0002g) in 250mL Erlenmeyer flask, add 100mL distilled water, 60 DEG C of water-bath 30min, get 40mL supernatant liquor, add 2 ~ 3 phenolphthalein indicators, be titrated to pink with standard NaOH solution, 30s is colour-fast.
Lactic acid (in total acid, %)=(V-V 0) × C × 0.09008 × 100/ (M × 40/100)
In formula: V-sample consumes the volume of NaOH, mL;
V 0-blank the volume consuming NaOH, mL;
0.09008-every milligramequivalent NaOH is equivalent to the grams of lactic acid;
M-sample quality, g.
PH measures: take 5g fermentation cotton dregs dregs of rice sample and be placed in 100mL triangular flask, add 45mL distilled water, shake up rear standing 30min, use determination of electrode pH.
1.5, data processing
Data acquisition SPSS16.0 software carries out one-way analysis of variance, adopts Duncan method to carry out multiple comparisons.Measurement result represents with " mean+SD ".
2, results and analysis
The impact that the chemical composition of table 1 cotton dregs raw material and moisture addition ferment on cotton dregs
Group Crude protein (%) Little peptide (accounting for crude protein, %) Lactic acid (in total acid, %) pH
Raw material 49.32±0.25 c 5.00±0.31 d 1.24±0.13 e 6.35
T 1 51.07±0.47 ab 15.23±0.47 c 3.60±0.02 b 5.40
T 2 51.68±0.26 a 16.27±0.13 a 3.67±0.04 a 5.34
T 3 50.46±0.12 b 15.34±0.40 bc 3.54±0.03 c 5.47
T 4 50.60±0.23 b 15.65±0.42 b 3.58±0.02 bc 5.43
T 5 50.35±0.38 b 15.82±0.24 b 3.21±0.05 d 5.71
Note: same column shoulder mark is different represents significant difference (p<0.05)
As shown in Table 1, after cotton dregs fermentation, the crude protein of all fermentation groups, little peptide and lactic acid content all significantly improve (P<0.05) than the cotton dregs that do not ferment, and pH obviously reduces.Wherein T 2the gross protein value of group (namely adding the moisture of 50% in fermention medium) reaches 51.68%, is significantly higher than T 3, T 4, T 5three groups, a little more than T 1group; Little peptide content, up to 16.27% (accounting for crude protein), is significantly higher than T 1, T 3, T 4, T 5four groups; Lactic acid content reaches 3.67, is significantly higher than T 1, T 3, T 4, T 5four groups.Comprehensive each Indexes Comparison, finds T 2group ferment effect is better than other five groups, and namely the ferment effect of moisture to cotton dregs of interpolation 50% is best.
The impact that embodiment 2 fermentation time ferments on cotton dregs
1, materials and methods
1.1, bacterial classification
Test composite bacteria used to be made up of plant lactobacillus, subtilis, Bacillus licheniformis, yeast saccharomyces cerevisiae, the ratio of above-mentioned four kinds of bacterium is 3:2:2:2.
1.2, cotton dregs raw material sources
Cotton dregs, purchased from China Oil and Food Import and Export Corporation (Changji) cereal and oil industry company limited, are that the face cake of cottonseed after squeezing is separated through most of Residual oil of extract technology by the inside again, a kind of by product obtained.
1.3, test design
Take cotton dregs 200g, according to the method for embodiment 1, in adjustment fermentation raw material, moisture is 50%, leavening temperature controls at 37 DEG C, fermentation time is respectively 1d, 2d, 3d, 4d, 5d, 6d, 80 DEG C of oven dry after fermentation ends, cross 40 mesh sieves after crushed, and measure the indexs such as crude protein, little peptide, lactic acid.
1.4, chemical analysis
A, gross protein value detect: perform by GB/T5009.5-2003 regulation.
B, little peptide content detect and adopt trichoroacetic acid(TCA) extraction method: accurately take fermentation cotton dregs sample 1.5g (being accurate to 0.0001g) in 100mL beaker, add 15% solution of trichloroacetic acid 25mL, vibration 5min, leave standstill 5min, solution is quantitatively shifted, at 3,000 rpm centrifugal 10min, get whole supernatant liquid filtering, accurately pipette 10mL filtrate and be placed in alimentary canal, digest by GB/T5009.5-2003 method and measure protein content.Little peptide calculation formula is as follows:
Little peptide (accounting for crude protein, %)=(V-V 0) × C × 6.25 × 0.014 × 2.5 × 100/ (M × CP)
In formula: V-sample consumes the volume of HCl, mL;
V 0-blank the volume consuming HCl, mL;
The volumetric molar concentration of C-hydrochloric acid, mol/L;
M-sample quality, g;
6.25 × 0.014-albumen gain factor;
The gross protein value of CP-fermentation cotton dregs sample.
C, lactic acid content (in total acid) adopt NaOH titration measuring: take 0.5g and ferment cotton dregs sample (being accurate to 0.0002g) in 250mL Erlenmeyer flask, add 100mL distilled water, 60 DEG C of water-bath 30min, get 40mL supernatant liquor, add 2 ~ 3 phenolphthalein indicators, be titrated to pink with standard NaOH solution, 30s is colour-fast.
Lactic acid (in total acid, %)=(V-V 0) × C × 0.09008 × 100/ (M × 40/100)
In formula: V-sample consumes the volume of NaOH, mL;
V 0-blank the volume consuming NaOH, mL;
0.09008-every milligramequivalent NaOH is equivalent to the grams of lactic acid;
M-sample quality, g.
PH measures: take 5g fermentation cotton dregs dregs of rice sample and be placed in 100mL triangular flask, add 45mL distilled water, shake up rear standing 30min, use determination of electrode pH.
1.5, data processing
Data acquisition SPSS16.0 software carries out one-way analysis of variance, adopts Duncan method to carry out multiple comparisons.Measurement result represents with " mean+SD ".
2, results and analysis
The impact that the table 2 different fermentations time ferments on cotton dregs
As shown in Table 2, along with the prolongation of fermentation time, the crude protein of fermentation cotton dregs, little peptide and lactic acid content all progressively increase, pH progressively reduces, and after fermentation 4d, rising tendency is comparatively mild, continue to extend fermentation time to 5d or 6d, gross protein value does not significantly increase.Consider that fermentation period prolongation can increase the problems such as production cost, preferred fermentation time is 4d.
The single bacterium of embodiment 3 and compound bacteria-fermented simultaneous test
1, materials and methods
1.1, bacterial classification
Single bacterium fermentation test adopts bacterial classification to be respectively plant lactobacillus, yeast saccharomyces cerevisiae, subtilis, Bacillus licheniformis, and after liquid nutrient medium is cultivated, gained bacterial classification concentration is 0.5 × 10 8~ 2 × 10 8cFU/mL; Compound bacteria-fermented is tested composite bacteria used and is made up of plant lactobacillus, subtilis, Bacillus licheniformis, yeast saccharomyces cerevisiae, and the ratio of four kinds of bacterium is 3:2:2:2.
1.2, cotton dregs raw material sources
Cotton dregs, purchased from China Oil and Food Import and Export Corporation (Changji) cereal and oil industry company limited, are that the face cake of cottonseed after squeezing is separated through most of Residual oil of extract technology by the inside again, a kind of by product obtained.
1.3, test design
Take cotton dregs 200g, according to the method for embodiment 1, in adjustment fermentation raw material, moisture is 50%, and single bacterium or composite bacteria add-on are 1 × 10 8cFU, leavening temperature controls at 37 DEG C, and fermentation time is 4d, 80 DEG C of oven dry after fermentation ends, crosses 40 mesh sieves after crushed, and measures the indexs such as crude protein, little peptide, lactic acid.
1.4, chemical analysis
A, gross protein value detect: perform by GB/T5009.5-2003 regulation.
B, little peptide content detect and adopt trichoroacetic acid(TCA) extraction method: accurately take fermentation cotton dregs sample 1.5g (being accurate to 0.0001g) in 100mL beaker, add 15% solution of trichloroacetic acid 25mL, vibration 5min, leave standstill 5min, solution is quantitatively shifted, at 3,000 rpm centrifugal 10min, get whole supernatant liquid filtering, accurately pipette 10mL filtrate and be placed in alimentary canal, digest by GB/T5009.5-2003 method and measure protein content.Little peptide calculation formula is as follows:
Little peptide (accounting for crude protein, %)=(V-V 0) × C × 6.25 × 0.014 × 2.5 × 100/ (M × CP)
In formula: V-sample consumes the volume of HCl, mL;
V 0-blank the volume consuming HCl, mL;
The volumetric molar concentration of C-hydrochloric acid, mol/L;
M-sample quality, g;
6.25 × 0.014-albumen gain factor;
The gross protein value of CP-fermentation cotton dregs sample.
C, lactic acid content (in total acid) adopt NaOH titration measuring: take 0.5g and ferment cotton dregs sample (being accurate to 0.0002g) in 250mL Erlenmeyer flask, add 100mL distilled water, 60 DEG C of water-bath 30min, get 40mL supernatant liquor, add 2 ~ 3 phenolphthalein indicators, be titrated to pink with standard NaOH solution, 30s is colour-fast.
Lactic acid (in total acid, %)=(V-V 0) × C × 0.09008 × 100/ (M × 40/100)
In formula: V-sample consumes the volume of NaOH, mL;
V 0-blank the volume consuming NaOH, mL;
0.09008-every milligramequivalent NaOH is equivalent to the grams of lactic acid;
M-sample quality, g.
PH measures: take 5g fermentation cotton dregs dregs of rice sample and be placed in 100mL triangular flask, add 45mL distilled water, shake up rear standing 30min, use determination of electrode pH.
1.5, data processing
Data acquisition SPSS16.0 software carries out one-way analysis of variance, adopts Duncan method to carry out multiple comparisons.Measurement result represents with " mean+SD ".
2, results and analysis
The impact that the single bacterium of table 3 and composite bacteria are fermented on cotton dregs
As shown in Table 3, crude protein, the little peptide of compound bacteria-fermented cotton dregs (account for crude protein, %) and lactic acid content be all significantly higher than the cotton dregs that plant lactobacillus, yeast saccharomyces cerevisiae, subtilis and Bacillus licheniformis ferment separately, and pH value is lower than single bacterium fermentation cotton dregs, be easy to preserve.
The single bacterium of embodiment 4 and compound bacteria-fermented simultaneous test
1, materials and methods
1.1, bacterial classification
Single bacterium fermentation test adopts bacterial classification to be respectively plant lactobacillus, yeast saccharomyces cerevisiae, subtilis, Bacillus licheniformis, and after liquid nutrient medium is cultivated, gained bacterial classification concentration is 0.5 × 10 8~ 2 × 10 8cFU/mL; Compound bacteria-fermented is tested composite bacteria used and is made up of plant lactobacillus, subtilis, Bacillus licheniformis, yeast saccharomyces cerevisiae, and the ratio of four kinds of bacterium is 1:3:1:3.
1.2, cotton dregs raw material sources
Cotton dregs, purchased from China Oil and Food Import and Export Corporation (Changji) cereal and oil industry company limited, are that the face cake of cottonseed after squeezing is separated through most of Residual oil of extract technology by the inside again, a kind of by product obtained.
1.3, test design
Take cotton dregs 200g, according to the method for embodiment 1, in adjustment fermentation raw material, moisture is 50%, and single bacterium or composite bacteria add-on are 1 × 10 8cFU, leavening temperature controls at 37 DEG C, and fermentation time is 4d, 80 DEG C of oven dry after fermentation ends, crosses 40 mesh sieves after crushed, and measures the indexs such as crude protein, little peptide, lactic acid.
1.4, chemical analysis
A, gross protein value detect: perform by GB/T5009.5-2003 regulation.
B, little peptide content detect and adopt trichoroacetic acid(TCA) extraction method: accurately take fermentation cotton dregs sample 1.5g (being accurate to 0.0001g) in 100mL beaker, add 15% solution of trichloroacetic acid 25mL, vibration 5min, leave standstill 5min, solution is quantitatively shifted, at 3,000 rpm centrifugal 10min, get whole supernatant liquid filtering, accurately pipette 10mL filtrate and be placed in alimentary canal, digest by GB/T5009.5-2003 method and measure protein content.Little peptide calculation formula is as follows:
Little peptide (accounting for crude protein, %)=(V-V 0) × C × 6.25 × 0.014 × 2.5 × 100/ (M × CP)
In formula: V-sample consumes the volume of HCl, mL;
V 0-blank the volume consuming HCl, mL;
The volumetric molar concentration of C-hydrochloric acid, mol/L;
M-sample quality, g;
6.25 × 0.014-albumen gain factor;
The gross protein value of CP-fermentation cotton dregs sample.
C, lactic acid content (in total acid) adopt NaOH titration measuring: take 0.5g and ferment cotton dregs sample (being accurate to 0.0002g) in 250mL Erlenmeyer flask, add 100mL distilled water, 60 DEG C of water-bath 30min, get 40mL supernatant liquor, add 2 ~ 3 phenolphthalein indicators, be titrated to pink with standard NaOH solution, 30s is colour-fast.
Lactic acid (in total acid, %)=(V-V 0) × C × 0.09008 × 100/ (M × 40/100)
In formula: V-sample consumes the volume of NaOH, mL;
V 0-blank the volume consuming NaOH, mL;
0.09008-every milligramequivalent NaOH is equivalent to the grams of lactic acid;
M-sample quality, g.
PH measures: take 5g fermentation cotton dregs dregs of rice sample and be placed in 100mL triangular flask, add 45mL distilled water, shake up rear standing 30min, use determination of electrode pH.
1.5, data processing
Data acquisition SPSS16.0 software carries out one-way analysis of variance, adopts Duncan method to carry out multiple comparisons.Measurement result represents with " mean+SD ".
2, results and analysis
The impact that the single bacterium of table 4 and composite bacteria are fermented on cotton dregs
As shown in Table 4, crude protein, the little peptide of compound bacteria-fermented cotton dregs (account for crude protein, %) and lactic acid content be all significantly higher than the cotton dregs that plant lactobacillus, yeast saccharomyces cerevisiae, subtilis and Bacillus licheniformis ferment separately, and pH value is lower than single bacterium fermentation cotton dregs, be easy to preserve.
The single bacterium of embodiment 5 and compound bacteria-fermented simultaneous test
1, materials and methods
1.1, bacterial classification
Single bacterium fermentation test adopts bacterial classification to be respectively plant lactobacillus, yeast saccharomyces cerevisiae, subtilis, Bacillus licheniformis, and after liquid nutrient medium is cultivated, gained bacterial classification concentration is 0.5 × 10 8~ 2 × 10 8cFU/mL; Compound bacteria-fermented is tested composite bacteria used and is made up of plant lactobacillus, subtilis, Bacillus licheniformis, yeast saccharomyces cerevisiae, and the ratio of four kinds of bacterium is 5:1:3:1.
1.2, cotton dregs raw material sources
Cotton dregs, purchased from China Oil and Food Import and Export Corporation (Changji) cereal and oil industry company limited, are that the face cake of cottonseed after squeezing is separated through most of Residual oil of extract technology by the inside again, a kind of by product obtained.
1.3, test design
Take cotton dregs 200g, according to the method for embodiment 1, in adjustment fermentation raw material, moisture is 50%, and single bacterium or composite bacteria add-on are 1 × 10 8cFU, leavening temperature controls at 37 DEG C, and fermentation time is 4d, 80 DEG C of oven dry after fermentation ends, crosses 40 mesh sieves after crushed, and measures the indexs such as crude protein, little peptide, lactic acid.
1.4, chemical analysis
A, gross protein value detect: perform by GB/T5009.5-2003 regulation.
B, little peptide content detect and adopt trichoroacetic acid(TCA) extraction method: accurately take fermentation cotton dregs sample 1.5g (being accurate to 0.0001g) in 100mL beaker, add 15% solution of trichloroacetic acid 25mL, vibration 5min, leave standstill 5min, solution is quantitatively shifted, at 3,000 rpm centrifugal 10min, get whole supernatant liquid filtering, accurately pipette 10mL filtrate and be placed in alimentary canal, digest by GB/T5009.5-2003 method and measure protein content.Little peptide calculation formula is as follows:
Little peptide (accounting for crude protein, %)=(V-V 0) × C × 6.25 × 0.014 × 2.5 × 100/ (M × CP)
In formula: V-sample consumes the volume of HCl, mL;
V 0-blank the volume consuming HCl, mL;
The volumetric molar concentration of C-hydrochloric acid, mol/L;
M-sample quality, g;
6.25 × 0.014-albumen gain factor;
The gross protein value of CP-fermentation cotton dregs sample.
C, lactic acid content (in total acid) adopt NaOH titration measuring: take 0.5g and ferment cotton dregs sample (being accurate to 0.0002g) in 250mL Erlenmeyer flask, add 100mL distilled water, 60 DEG C of water-bath 30min, get 40mL supernatant liquor, add 2 ~ 3 phenolphthalein indicators, be titrated to pink with standard NaOH solution, 30s is colour-fast.
Lactic acid (in total acid, %)=(V-V 0) × C × 0.09008 × 100/ (M × 40/100)
In formula: V-sample consumes the volume of NaOH, mL;
V 0-blank the volume consuming NaOH, mL;
0.09008-every milligramequivalent NaOH is equivalent to the grams of lactic acid;
M-sample quality, g.
PH measures: take 5g fermentation cotton dregs dregs of rice sample and be placed in 100mL triangular flask, add 45mL distilled water, shake up rear standing 30min, use determination of electrode pH.
1.5, data processing
Data acquisition SPSS16.0 software carries out one-way analysis of variance, adopts Duncan method to carry out multiple comparisons.Measurement result represents with " mean+SD ".
2, results and analysis
The impact that the single bacterium of table 5 and composite bacteria are fermented on cotton dregs
As shown in Table 5, crude protein, the little peptide of compound bacteria-fermented cotton dregs (account for crude protein, %) and lactic acid content be all significantly higher than the cotton dregs that plant lactobacillus, yeast saccharomyces cerevisiae, subtilis and Bacillus licheniformis ferment separately, and pH value is lower than single bacterium fermentation cotton dregs, be easy to preserve.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (10)

1. a composite bacteria, is characterized in that, comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
2. composite bacteria according to claim 1, it is characterized in that, in CFU, the consumption of described plant lactobacillus, described subtilis, described Bacillus licheniformis and described yeast saccharomyces cerevisiae is (1 ~ 5): (1 ~ 3): (1 ~ 3): (1 ~ 3).
3. composite bacteria according to claim 2, is characterized in that, in CFU, the consumption of described plant lactobacillus, described subtilis, described Bacillus licheniformis and described yeast saccharomyces cerevisiae is 3:2:2:2.
4. a cotton dregs fermented feed, is characterized in that, its preparation method comprises:
Cotton dregs are mixed with water, obtains fermentation material, described fermentation material is mixed with composite bacteria, adopt solid state fermentation to ferment, dry, obtain cotton dregs fermented feed;
Described composite bacteria comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
5. cotton dregs fermented feed according to claim 4, is characterized in that, the mass percentage of described water in described fermentation material is 45% ~ 65%.
6. cotton dregs fermented feed according to claim 4, is characterized in that, in CFU/g, the consumption of described composite bacteria and described cotton dregs is (2.5 ~ 10) × 10 8: 1000.
7. cotton dregs fermented feed according to claim 4, is characterized in that, the temperature of described solid state fermentation is 36 ~ 38 DEG C.
8. cotton dregs fermented feed according to claim 4, is characterized in that, the time of described solid state fermentation is 1 ~ 6d.
9. cotton dregs fermented feed according to claim 4, is characterized in that, the temperature of described oven dry is 70 ~ 90 DEG C.
10. a preparation method for cotton dregs fermented feed, is characterized in that, comprises the steps:
Cotton dregs are mixed with water, obtains fermentation material, described fermentation material is mixed with composite bacteria, adopt solid state fermentation to ferment, dry, obtain cotton dregs fermented feed;
Described composite bacteria comprises plant lactobacillus, subtilis, Bacillus licheniformis and yeast saccharomyces cerevisiae.
CN201511021332.9A 2015-12-30 2015-12-30 Compound bacteria/cottonseed meal fermented feed and preparation method thereof Pending CN105543131A (en)

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CN110236002A (en) * 2019-07-18 2019-09-17 山东鸿盛牧场生物科技有限公司 A kind of cattle and sheep fermented feed and preparation method thereof

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