CN111685240A - Research method of waterfowl biological fermentation feed - Google Patents
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- 238000011160 research Methods 0.000 title claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, 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
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
-
- Y—GENERAL 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention provides a research method of waterfowl biological fermentation feed, and relates to the technical field of waterfowl biological fermentation feed. The research method of the waterfowl biological fermented feed comprises the steps of fermented feed proportioning, flora acquisition, growth characteristic measurement of each bacterium, determination of mixed feed fermentation conditions and Plackett-Burman test design. Through the culture and screening of flora and the comprehensive analysis of different experiments by utilizing a Plackett-Burman tool, the formula of PH, water content and temperature is obtained under the condition that the feed ratio and the ratio of each bacterium are determined, and scientific basis is provided for the actual production.
Description
Technical Field
The invention relates to the technical field of waterfowl biological fermentation feed, in particular to a research method of waterfowl biological fermentation feed.
Background
The feed prepared by fermenting the coarse feed by microorganisms is fermented feed, the coarse feed is rich in coarse fibers such as cellulose, hemicellulose, pectin substances, lignin and the like and protein, but is difficult to be directly digested and absorbed by animals, and the eating of the coarse feed can increase the intestinal burden and cause intestinal diseases.
Aiming at the problems of culture environment pollution in waterfowl production, feed antibiotics to be forbidden in culture and the like, the development of the waterfowl biological fermentation feed has wide prospect.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a research method of a waterfowl biological fermentation feed, which solves the problems of culture environment pollution in waterfowl production and feed antibiotics to be forbidden in culture.
(II) technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme: a research method of waterfowl biological fermentation feed comprises the following steps:
s1 fermented feed proportioning
According to the nutritional requirements of ducks, by combining the development and utilization of unconventional feed raw materials, the fermented feed raw materials are determined to be 48% of corn, 12% of bran, 32% of bean pulp and 8% of rapeseed meal;
s2 acquisition of flora
The bacillus and the lactobacillus are primarily screened from chicken manure, pig manure, duck manure, goose manure, fermented soybean meal, fermented feed and yoghourt, are selected according to the ratio of the calcium-solubilizing rings to the strains with large diameter, are inoculated for ten times, 20 strains from different sources are obtained by observing stability, and the yeast is obtained by culturing from high-activity dry yeast powder;
s3 determination of growth characteristics of various bacteria
In order to obtain the optimal seed age of each strain finally used for mixed feed fermentation, the growth characteristics of each strain are measured;
s4 determination of fermentation conditions of mixed feed
Obtaining the optimal feed fermentation condition by carrying out batch gradient experiments on different water contents, different inoculation amounts and different proportions of strains;
s5 Plackett-Burman test design
By analyzing the data obtained in S4, the equation of PH, water content and temperature is obtained by using acid soluble protein as an index, thereby providing a basis for actual production.
(III) advantageous effects
The invention provides a research method of waterfowl biological fermentation feed. The method has the following beneficial effects:
1. according to the invention, the bacillus and the lactobacillus are indirectly screened and cultured from livestock, and the optimal fermentation data of the feed is obtained through various experiments, so that the pollution of the culture environment of waterfowls in production can be effectively reduced, and the problem of feed antibiotics to be forbidden in culture can be avoided.
2. According to the invention, through the comprehensive analysis of the Plackett-Burman tool on different experiments, the equation of PH, water content and temperature is obtained under the condition that the feed ratio and the ratio of each bacterium are determined, so that scientific basis is provided for actual production, the quality of the fermented feed is improved, and the production cost of the fermented feed is reduced.
Drawings
FIG. 1 is a schematic diagram of the growth curve of each strain of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
as shown in figure 1, a research method of waterfowl biological fermentation feed comprises the following steps:
s1 fermented feed proportioning
According to the nutritional requirements of ducks, by combining the development and utilization of unconventional feed raw materials, the fermented feed raw materials are determined to be 48% of corn, 12% of bran, 32% of bean pulp and 8% of rapeseed meal;
s2 acquisition of flora
2.1: primary and secondary screening of bacillus
Primarily screening chicken manure, pig manure, goose manure, natto, fermented soybean meal and fermented feed, selecting according to the large diameter of a lysoprotein ring, performing ten times of inoculation, and observing stability to obtain 20 strains from different sources.
And (3) selecting the primarily screened bacillus strains again, selecting 4 strains from each animal and plant, and measuring neutral protein, acid protein, cellulase and a-amylase. Selecting strains with high enzyme activity according to the measured value, carrying out streak culture, selecting single colony, and measuring with high temperature resistance (40 deg.C, 50 deg.C, 60 deg.C) and acid resistance (PH 3, 4, 5, 6, 7). The screened bacillus is a chicken hard 3 strain, and specific results are shown in tables 1 and 2;
TABLE 1 determination of the enzymatic Activity of Bacillus
TABLE 2 acid and high temperature resistance assay of Bacillus
2.2: primary and secondary screening of lactobacillus
The method comprises the steps of primary screening from chicken manure, pig manure, duck manure, goose manure, fermented soybean meal, fermented feed and yoghourt, performing ten times of inoculation according to the selection of a calcium-dissolving ring and a strain with a large diameter ratio, and observing stability to obtain 20 strains from different sources.
And (3) selecting 4 bacteria from each source for streak culture according to the large ratio of the diameter of the calcium dissolving ring to the diameter of the bacteria, selecting a single bacterial colony for liquid culture, and performing pH determination after 48h of culture. The strains screened in this experiment were derived from the plant lactobacillus yoghurt 3, the results are detailed in table 3;
TABLE 3 determination of the pH of lactic acid bacteria from different sources
2.3: yeast
Culturing the yeast from the high-activity dry yeast powder to obtain yeast;
s3 determination of growth characteristics of various bacteria
In order to obtain the optimum seed age of each strain to be finally used for the mixed feed fermentation, the growth characteristics of each strain were measured. And (3) carrying out streak culture on the bacillus (chicken hard 3), the lactic acid bacteria (yoghourt 3) and the yeast left after re-screening, and selecting a single colony for liquid culture. Firstly, 100ml of seed liquid culture medium, MRS liquid culture medium and potato liquid culture medium are prepared, 5ml of seed liquid culture medium, MRS liquid culture medium and potato liquid culture medium are respectively taken and subpackaged into test tubes, a proper amount of single strains of bacillus, lactobacillus plantarum and saccharomycetes are selected and inoculated into the subpackaged test tubes, the test tubes are uniformly shaken by oscillation, the lactobacillus plantarum is shaken in an oscillator for 12 hours at 37 ℃ and 28 ℃, and the lactobacillus plantarum is placed in an incubator for 12 hours at 37 ℃ under anaerobic conditions. The separated test tubes are marked for 0h, 2h, 4h, 6h, 8h, 10h, 12h, 14h, 16h, 18h, 20h, 22h, 24h and 26 h. Respectively sucking 50 μ l of the cultured bacteria liquid into labeled test tubes, placing bacillus in an oscillator at 37 deg.C and yeast at 28 deg.C, standing lactobacillus plantarum in an incubator at 37 deg.C under anaerobic condition, and respectively taking out and measuring absorbance value according to time. The growth curve was plotted with time as the horizontal coordinate and absorbance as the vertical coordinate.
As shown in figure 1, the bacillus grows rapidly in 0-10h, the bacillus grows negatively in 10-12h, grows rapidly in 12-20h, and after 20h, the bacillus enters a growth and death period, and the number of the bacillus reaches the maximum value after 20 h. The lactobacillus is in the growth delay period of 0-2h, the lactobacillus is in the rapid growth period of 2-12h, the lactobacillus enters the growth decay period of 12h, and the lactobacillus count is in the maximum value of 12 h. 0-2h of yeast is the growth lag phase of the thallus, 2-8h is the rapid growth phase of the thallus, 8-10h of the thallus growth speed becomes slow, 10-12h of the thallus growth speed returns, and 12h is the maximum value of the thallus quantity;
s4 determination of fermentation conditions of mixed feed
4.1: influence of different water contents on fermentation effect of mixed feed
Adjusting the pH value of the mixed feed (consisting of corn, bran, bean pulp and rapeseed meal) to 7.02, subpackaging the mixed feed into 5 sealed bags with the same mass, adding 10 percent (20ml) of bacterial liquid into all the bags according to the experimental design, respectively adding 40 percent (60ml), 50 percent (80ml), 60 percent (100ml), 70 percent (120ml) and 80 percent (140ml) of ultrapure water into the bags, uniformly stirring, sealing, and culturing in a constant-temperature incubator at 35 ℃ for 3 days. The above-mentioned bacterial liquid is the bacterial liquid obtained by culturing the above-mentioned bacillus, lactic acid bacteria and saccharomycetes for 24 hr. Sampling 24h, 48h and 72h after fermentation to determine a pH value, acid soluble protein and reducing sugar;
TABLE 4 Effect of different water contents on the fermentation Effect of Bacillus
TABLE 5 Effect of different Water contents on the fermentation Effect of lactic acid bacteria
TABLE 6 influence of different water contents on the fermentation effect of yeasts
4.2: influence of different inoculation amounts on fermentation effect of mixed feed
Adjusting the pH value of the mixed feed to 7.02, subpackaging the mixed feed into 6 sealed bags with the same mass, adding 10ml of bacterial liquid and 110ml of ultrapure water into a 5% inoculation amount bag according to experimental design, adding 20ml of bacterial liquid and 100ml of ultrapure water into a 10% inoculation amount bag, adding 30ml of bacterial liquid and 90ml of ultrapure water into a 15% inoculation amount bag, adding 40ml of bacterial liquid and 80ml of ultrapure water into a 20% inoculation amount bag, adding 50ml of bacterial liquid and 70ml of ultrapure water into a 25% inoculation amount bag, adding 60ml of bacterial liquid and 60ml of ultrapure water into a 30% inoculation amount bag, uniformly stirring, sealing and placing the mixture in a 35 ℃ constant temperature incubator for fermentation for 3 days. The water content of the feed inoculated with each inoculation amount is the same and is 60 percent. Sampling and measuring a ph value, acid soluble protein and reducing sugar 24h, 48h and 72h after fermentation;
TABLE 7 influence of different inoculum sizes on the fermentation efficiency of Bacillus
TABLE 8 influence of different inoculum sizes on the fermentation effect of yeast lactic acid bacteria
TABLE 9 influence of different inoculum sizes on the fermentation effect of yeasts
4.3: influence of different proportions of strains on fermentation effect of mixed feed
And (3) carrying out streak culture on various strains left after re-screening, selecting a single colony to carry out liquid culture (the lactobacillus phytate is cultured for 24 hours, and the yeast and the bacillus are cultured for 12 hours), centrifuging the cultured bacterial liquid (3500r/min is converted into 8min), pouring out the culture liquid, carrying out centrifugal washing twice by using normal saline, and continuously adding the normal saline to shake and mix uniformly. Adjusting the pH value of the mixed feed to 7.02 (using dilute sodium hydroxide), subpackaging the samples into 9 sealed bags with the same mass, distributing the strain inoculation amount of 10% in each bag according to the experimental design (orthogonal design) but distributing the proportion of the three strains according to the proportion in the table 10, adding the strain liquid and ultrapure water, uniformly stirring, sealing, and placing in a 35 ℃ constant temperature incubator for fermentation for 3 days. Sampling 72h after fermentation to determine the content of acid soluble protein. The results are shown in Table 11;
TABLE 10 proportion of three bacteria in the fermentation process
TABLE 11 influence of different ratios of strains on the fermentation results
According to the water content, the inoculation amount and the strain proportion test, the optimum water content of the biological fermentation feed is determined to be 50%, the inoculation amount is determined to be 10%, and the ratio of bacillus: yeast: the ratio of lactic acid bacteria is 3:1: 2;
s5 Plackett-Burman test design
In order to determine the most key influencing factors influencing the fermentation effect, different gradients are set for factors such as temperature, moisture, inoculum size, pH, strain ratio and the like, and the results show that the water content, the pH and the temperature are the main factors influencing the fermentation effect. The specific results are shown in tables 12 and 13;
TABLE 12Plackett-Burman test design and response values
TABLE 13 Plackett-Burman test design regression analysis results
According to the fact that the water content, the pH and the temperature are determined to be main factors influencing the fermentation effect, the optimal water content, the pH and the temperature are determined through a response surface test. The optimal reaction conditions obtained by using acid soluble protein as a main index and reducing sugar as an auxiliary index are that the pH is 7.200, the water content is 50.235 percent, and the culture temperature is 40 ℃. And the equations of acid soluble protein, water content, pH and temperature are given as:
acid soluble protein-2.38952 +0.30487 ph +0.029587 water content +0.13388 temperature.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (1)
1. A research method of waterfowl biological fermentation feed comprises the following steps:
s1 fermented feed proportioning
According to the nutritional requirements of ducks, by combining the development and utilization of unconventional feed raw materials, the fermented feed raw materials are determined to be 48% of corn, 12% of bran, 32% of bean pulp and 8% of rapeseed meal;
s2 acquisition of flora
Primarily screening chicken manure, pig manure, duck manure, goose manure, fermented soybean meal, fermented feed and yoghourt, selecting according to the ratio of calcium-dissolving rings to strains is large, inoculating for ten times, observing stability to obtain 20 strains from different sources, and culturing yeast from high-activity dry yeast powder;
s3 determination of growth characteristics of various bacteria
In order to obtain the optimal seed age of each strain finally used for mixed feed fermentation, the growth characteristics of each strain are measured;
s4 determination of fermentation conditions of mixed feed
Obtaining the optimal feed fermentation condition by carrying out batch gradient experiments on different water contents, different inoculation amounts and different proportions of strains;
s5 Plackett-Burman test design
By analyzing the data obtained by S4, the equation of PH, water content and temperature is obtained by taking acid soluble protein as an index, thereby providing scientific basis for actual production.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039699A (en) * | 2013-01-05 | 2013-04-17 | 湖南省微生物研究所 | Mixed fermentation feed and preparation method and application method of mixed fermentation feed |
| CN107319172A (en) * | 2017-07-06 | 2017-11-07 | 江苏农牧科技职业学院 | A kind of family's poultry micro-ecological preparation and preparation method thereof |
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2020
- 2020-04-21 CN CN202010315749.0A patent/CN111685240A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039699A (en) * | 2013-01-05 | 2013-04-17 | 湖南省微生物研究所 | Mixed fermentation feed and preparation method and application method of mixed fermentation feed |
| CN107319172A (en) * | 2017-07-06 | 2017-11-07 | 江苏农牧科技职业学院 | A kind of family's poultry micro-ecological preparation and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 王哲奇等: ""响应面优化混菌固态发酵豆粕制备多肽饲料的发酵条件"", 《饲料工业》 * |
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Application publication date: 20200922 |
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