CN115039833A - Preparation method of yellow-stored straw feed by-10 ℃ non-freezing bacterium-enzyme synergistic fermentation - Google Patents
Preparation method of yellow-stored straw feed by-10 ℃ non-freezing bacterium-enzyme synergistic fermentation Download PDFInfo
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- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 12
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- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 12
- 241001468157 Lactobacillus johnsonii Species 0.000 claims abstract description 12
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- 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
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- 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
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Abstract
A preparation method of yellow-stored straw feed by bacterium-enzyme synergistic fermentation at minus 10 ℃ without freezing relates to a preparation method of yellow-stored feed, and aims to solve the technical problem that the yellow-stored feed is frozen at a low temperature below zero in winter so as to influence the fermentation. The method comprises the following steps: uniformly mixing corn straws and a composite type antifreezing leavening agent to obtain a raw material; placing the raw materials in a silage bag for dark fermentation to obtain the compound antifreezing fermentation agent, wherein the compound antifreezing fermentation agent is prepared by uniformly mixing a microbial agent, active enzyme, a urea solution and sugar; the microbial agent is prepared by mixing bacillus subtilis subspecies, bacillus licheniformis, lactobacillus plantarum and lactobacillus johnsonii; the active enzyme is prepared by mixing cellulase, laccase, xylanase, pectinase and beta-glucanase; the mass percentage concentration of the urea solution is 30-35%. The yellow silage prepared by the method is not frozen and is fully fermented, and can be used in the field of winter feed preparation in cold regions.
Description
Technical Field
The invention relates to a yellow corn silage technology, in particular to a preparation technology of yellow corn silage straw feed by the synergistic fermentation of bacterium enzyme which does not freeze at the temperature of minus 10 ℃.
Background
Corn stalk is one of the dominant resources in northeast China, and the yield of the corn stalk accounts for about 48% of the total yield of China. However, due to the factors of insufficient technical means, unreasonable planning and the like, the comprehensive utilization rate of the corn straws is less than 70%. In recent years, with the continuous development of science and technology, the field of the five-purpose of the straws is continuously broken through, and the comprehensive utilization rate of the corn straws is stably improved. In particular, the requirements and demands for meat quality are also increasing due to the improvement of the living standard of people, and the ruminant meat can just meet the requirements as a high-quality meat food source. Therefore, this has also prompted the development of scale and intensification in ruminant breeding. The coarse fodder has irreplaceable special physiological needs for ruminants, and further reflects the urgent need of the development of the ruminant breeding industry for the coarse fodder, and the straw forage has the advantages of low cost, wide sources and large quantity, and is an important measure for developing the grass and livestock industry and improving the ecological environment.
The yellow corn silage technology is an important measure which can preserve nutrient substances of forage grass and can improve the digestibility, palatability and feed intake of the forage. But because the high cellulose contained in the straw limits the digestibility of animals, particularly the hard-to-dissociate mosaic structure formed by combining lignin with cellulose and hemicellulose. The ideal feeding effect is still difficult to achieve after simple yellow storage. Therefore, the low palatability and the digestion utilization rate of the yellow storage straw can be obviously improved by adding the ensiling microbial additive or the biological enzyme preparation which is specially used for cellulose degradation or lactic acid production and the like. However, in the northeast, harvesting and air-drying of corn stalks can be used in the late autumn and winter in the northeast, and the average temperature in the northeast is about-10 ℃ in winter, which causes the activity of microorganisms and biological enzymes to be greatly reduced, and finally the quality of yellow storage cannot be guaranteed. Furthermore, the breeding facilities and forage grass storage places of the herbivorous livestock such as cattle and sheep in northeast are open and heat-insulating facilities. Therefore, the yellow silage in northeast can be frozen into ice lumps in winter, feeding cannot be achieved at all, and meanwhile due to freezing, anaerobic fermentation microorganisms such as lactic acid bacteria and biological enzymes of the corn straws are difficult to play a role in the yellow silage process, so that the quality of the fermented feed is poor. If the problem of freezing of the yellow straw feed in winter can be solved, the problem of shortage of high-quality coarse feed of cattle and sheep in winter in northeast regions can be solved, and the growth performance of cattle and sheep in winter and the survival rate of lambing and calving can be improved.
Disclosure of Invention
The invention provides a preparation method of a yellow-stored straw feed by bacterium-enzyme synergistic fermentation at-10 ℃ in order to solve the technical problem that the yellow-stored feed is frozen in a low-temperature-below-zero state in winter and further influences fermentation. According to the invention, the microbial preparation and the biological enzyme preparation are added into the corn straws by adding the urea solution, and the urea solution is added, so that a feed nitrogen source is increased, freezing at the temperature of-10 ℃ can be avoided, and the optimal activity of the microbes and the enzymes under a low-temperature condition can be maintained, so that the fermentation quality of the yellow-stored feed is promoted, and the actual problem that the production and breeding are influenced by freezing of the low-temperature yellow-stored feed in winter is solved.
The preparation method of the-10 ℃ non-freezing fungus enzyme synergistic fermentation yellow straw feed comprises the following steps:
firstly, uniformly mixing corn straws and a composite antifreezing leavening agent to obtain a raw material; the composite antifreezing leaven is prepared from a microbial agent, an active enzyme, a urea solution and sugar in a mass ratio of (1-3): 1: (1-2): (1-2) uniformly mixing; the microbial agent is prepared from bacillus subtilis subspecies, bacillus licheniformis, lactobacillus plantarum and lactobacillus johnsonii according to the mass ratio of (1.5-2.4): 1: (3.2-4.5): (4.3-5.5) in proportion; the active enzyme is prepared from cellulase, laccase, xylanase, pectinase and beta-glucanase in a mass ratio of (1-3): (1-3): (1-2): 1: (1-2) mixing; the urea solution is 30-35% by mass;
and secondly, adjusting the water content of the raw materials obtained in the step one to be 50-65% by mass percent, compacting, placing the raw materials into a silage bag, vacuumizing and sealing the silage bag, and fermenting the silage bag in a dark place at the temperature of more than or equal to-10 ℃ for 30-45 days to obtain the yellow silage.
In a further step, the bacillus subtilis subspecies is purified bacillus subtilis subspecies GIM1.372 which is subjected to domestication and reversion verification at the low temperature of-15 ℃, and the strain is from Guangdong province microbial culture collection center (GDMCC).
Furthermore, the bacillus licheniformis is domesticated at a low temperature of-15 ℃, subjected to progenitor verification and purified bacillus licheniformis GIM1.182, and the strain is from Guangdong province microbial culture collection center (GDMCC).
Furthermore, the lactobacillus plantarum is subjected to domestication at a low temperature of-15 ℃, progenitor verification and purification, and the strain is from Guangdong province microbial culture collection center (GDMCC) 1.648.
Further, the lactobacillus johnsonii is subjected to domestication at a low temperature of-15 ℃, reversion verification and purification, and the lactobacillus johnsonii GIM1.730 is obtained from Guangdong province microbial culture collection center (GDMCC).
In a step, the corn stalks are cut into sections of 1cm to 3 cm.
And in the step one, the mass ratio of the composite antifreezing fermentation agent to the corn stalks in the step one is (0.2-0.6): 1000.
more preferably, the sugar in step one is sucrose.
According to the invention, a proper amount of urea solution is added and mixed with the microbial agent, the sugar and the active enzyme to prepare the compound type antifreezing fermentation agent, so that the problem that the activity of the microbial agent for corn straw yellow storage and the activity of the active enzyme are inhibited at a low-temperature state below zero is solved, the microorganisms can effectively decompose and utilize nutrient components in the straws to produce lactic acid, and the target effect of the active enzyme is matched to achieve the function of degrading lignin. The contents of acid washing fiber, neutral washing fiber, lignin and hemicellulose in the yellow storage feed produced by the method are further reduced, more nutrient components such as soluble sugar and lactic acid are generated, the in-vitro digestibility of dry matters, crude protein, neutral washing fiber and acid washing fiber of the yellow storage feed is also remarkably improved, and finally, the phenomenon that the yellow storage feed is frozen at the low temperature below zero in winter is effectively prevented.
According to the invention, through the bacterium-enzyme synergistic fermentation and urea ammoniation technologies, microorganisms and biological enzymes can still exert the maximum activity in an anaerobic environment under a low-temperature environment below zero, so that the yellow straw feed can not be frozen in winter, the straw digestibility and the nitrogen content are improved, the antifreezing effect is achieved, and the method can be applied to the field of winter feed preparation in cold regions.
Detailed Description
The following examples are used to demonstrate the beneficial effects of the present invention.
Example 1: the preparation method of the yellow straw feed comprises the following steps:
firstly, mixing bacillus subtilis subspecies, bacillus licheniformis, lactobacillus plantarum and lactobacillus johnsonii according to the mass ratio of 2: 1: 4.5: 5.3 uniformly mixing the bacillus subtilis subspecies, the bacillus licheniformis, the lactobacillus plantarum and the lactobacillus johnsonii to obtain a microbial agent; wherein the bacillus subtilis subspecies is the bacillus subtilis subspecies GIM1.372 which is purified after low-temperature domestication and reversion verification at-15 ℃; the bacillus licheniformis is domesticated at a low temperature of-15 ℃, subjected to progenitor verification and purified bacillus licheniformis GIM 1.182; the lactobacillus plantarum is lactobacillus plantarum GIM1.648 which is domesticated at a low temperature of-15 ℃, subjected to progenitor verification and purified; the lactobacillus johnsonii is subjected to domestication at a low temperature of-15 ℃, progenitor verification and purification, and is lactobacillus johnsonii GIM 1.730; bacillus subtilis subspecies subtilis GIM1.372, Bacillus licheniformis GIM1.182, Lactobacillus plantarum GIM1.648 and Lactobacillus johnsonii GIM1.730 are all from the Guangdong province collection of microorganisms (GDMCC);
then, according to the mass ratio of cellulase, laccase, xylanase, pectinase and beta-glucanase of 1: 1: 1: 1: 1, uniformly mixing cellulase, laccase, xylanase, pectinase and beta-glucanase to obtain active enzyme;
mixing a microbial agent, an active enzyme, a urea solution with the mass percentage concentration of 32% and sucrose according to the mass ratio of 1: 1: 1: 1, uniformly mixing to obtain a compound antifreezing leavening agent;
cutting the corn straws into sections of 2cm, and uniformly mixing the composite type antifreezing fermentation agent and the corn straws according to the mass ratio of the composite type antifreezing fermentation agent to the corn straws of 0.4:1000 to obtain a No. 1 raw material;
and secondly, adding water into the No. 1 raw material obtained in the step one, adjusting the water content to 55% by mass, then putting the mixture into an ensiling bag, vacuumizing the ensiling bag until the gauge pressure of a vacuum table is-0.02 MPa, sealing the ensiling bag, and fermenting the sealed ensiling bag in dark at the temperature of-10 ℃, 15 ℃ and 20 ℃ for 45 days to obtain yellow ensiling feed which is marked as C1-10, C1-15 and C1-20 feed.
Example 2: the difference between the embodiment and the embodiment 1 is that in the step one, the preparation method of the composite antifreezing fermentation agent comprises the following steps: mixing a microbial agent, an active enzyme, a urea solution with the mass percentage concentration of 32% and cane sugar according to the mass ratio of 2: 1: 1: 1, uniformly mixing to obtain a compound antifreezing leavening agent; the yellow stocks obtained were recorded as C2-10, C2-15 and C2-20 feeds in the same manner as in example 1.
Example 3: the difference between the embodiment and the embodiment 1 is that in the step one, the mass percentage concentration of the urea solution is 22%, the other steps are the same as the embodiment 1, and the obtained yellow silage is marked as C3-10, C3-15 and C3-20 feeds.
Example 4: the difference between the embodiment and the embodiment 2 is that in the step one, the mass percentage concentration of the urea solution is 22%, the yellow silage obtained by the embodiment 2 is marked as C4-10, C4-15 and C4-20 feeds.
Example 5: the difference between the embodiment and the embodiment 1 is that in the step one, the mass percentage concentration of the urea solution is 42%, and the yellow silage obtained by the embodiment 1 is marked as C5-10, C5-15 and C5-20 feeds.
Example 6: the difference between the embodiment and the embodiment 2 is that in the step one, the mass percentage concentration of the urea solution is 42%, the yellow silage obtained by the embodiment 2 is marked as C6-10, C6-15 and C6-20 feeds.
The six groups of feeds prepared in the embodiments 1-6 are subjected to apparent judgment to determine whether the feeds are frozen, and then the crude protein content, the pH value, the lactic acid content, the acidic detergent fiber content, the neutral detergent cellulose content and the acidic lignin content of the feeds are tested, and meanwhile, a proper amount of feed samples are taken to perform a rumen fluid in-vitro digestion test for 48 hours, and the test results are listed in table 1.
Table 1 results of performance test of each feed prepared in example 1
The results of the test were analyzed as follows:
after a compound leavening agent prepared by a microbial agent, active enzyme and a low-temperature anticoagulant of 22 percent urea solution or 32 percent urea solution or 42 percent urea solution is added at the temperature of minus 15 ℃ and minus 20 ℃, the phenomenon of icing occurs in yellow storage.
After a compound leavening agent prepared by a microbial agent, active enzyme and a urea solution with the concentration of 22 percent or a urea solution with the concentration of 42 percent is added at the temperature of minus 10 ℃, the phenomenon of icing occurs in yellow storage, and after the compound leavening agent prepared by the microbial agent, the active enzyme and a urea solution with the concentration of 32 percent is added, the phenomenon of icing does not occur in the yellow storage.
Therefore, the composite leaven prepared by the microbial agent, the active enzyme and the 32 percent urea solution can effectively solve the important problems that the freezing of the yellow storage feed at the temperature of minus 10 ℃ is serious and the breeding production is influenced.
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of-10 ℃ below zero: 1: 1: 1, uniformly mixing to prepare a C1-10 feed of the composite leavening agent, wherein the pH value is 4.56, and adding a microbial agent, an active enzyme, a 32% urea solution and sugar according to the mass ratio of 2: 1: 1: 1 the pH value of the C2-10 feed which is uniformly mixed to prepare the compound leaven is 4.04, which is obviously reduced by 0.52(P is less than 0.05) compared with the C1 feed.
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of-10 ℃ below zero: 1: 1: 1, uniformly mixing to prepare the compound leaven, wherein the content of neutral detergent fiber of the C1-10 feed is 56.44%, and adding a microbial agent, an active enzyme, a 32% urea solution and sugar according to a mass ratio of 2: 1: 1: 1 the content of neutral detergent fiber of the C2-10 feed which is uniformly mixed to prepare the compound leaven is 50.17 percent, and is obviously reduced by 6.27 percent compared with the C1-10 feed (P is less than 0.05).
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of-10 ℃ below zero: 1: 1: 1, uniformly mixing to prepare a composite leavening agent, wherein the content of C1-10 feed acidic detergent fiber is 46.48%, and adding a microbial agent, an active enzyme, a 32% urea solution and sugar according to the mass ratio of 2: 1: 1: 1 the content of the acid detergent fiber of the C2-10 feed which is uniformly mixed to prepare the compound leaven is 40.99 percent, which is obviously reduced by 5.49 percent compared with the C1-10 feed (P is less than 0.05).
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of 1: 1: 1: 1, uniformly mixing to prepare the composite leaven, wherein the content of C1-10 feed acidic washing lignin is 6.49%, and adding a microbial agent, an active enzyme, a 32% urea solution and sugar according to a mass ratio of 2: 1: 1: 1 the content of the acidic washing lignin of the C2-10 feed which is uniformly mixed to prepare the compound leaven is 3.79 percent, and is obviously reduced by 2.70 percent compared with the C1-10 feed (P is less than 0.05).
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of-10 ℃ below zero: 1: 1: 1, uniformly mixing to prepare the compound leaven, wherein the content of soluble sugar in the C1-10 feed is 11.07%, and adding a microbial agent, an active enzyme, a 32% urea solution and sugar according to a mass ratio of 2: 1: 1: 1 the soluble sugar content of the C2-10 feed prepared by uniformly mixing the components is 15.67 percent, and is obviously reduced by 4.60 percent (P is less than 0.05) compared with the C1-10 feed.
Adding a microbial agent, active enzyme, a 32% urea solution and sugar at the mass ratio of-10 ℃ below zero: 1: 1: 1 the content of lactic acid in the C1-10 feed prepared by uniformly mixing the components is only 2.07 percent, and the compound leaven has lower dry matter digestibility in vitro, crude protein digestibility in vitro, neutral detergent fiber digestibility in vitro and acid detergent fiber digestibility in vitro. Adding a microbial agent, active enzyme, a 32% urea solution and sugar according to a mass ratio of 2: 1: 1: 1 the lactic acid content of the C2-10 feed which is uniformly mixed to prepare the composite leaven can reach 3.23 percent, and the dry matter digestion rate in vitro, the crude protein digestion rate in vitro, the neutral detergent fiber digestion rate in vitro and the acid detergent fiber digestion rate in vitro are obviously improved.
This is because even at a temperature of-10 ℃, after the proper concentration of microbial agents is combined with sugar, active enzymes and 32% urea solution, the microorganisms can still maintain the maximum activity and play a greater functional role in a low-temperature and anaerobic state, and the sugar, the nutrients in the straws and the provided water are utilized to rapidly enter a fermentation mode, so that more lactic acid is generated, and the pH is significantly reduced. And the high-efficiency active enzyme is combined, so that the thick cell walls of the straws are more effectively degraded, the content of cellulose such as acid washing fiber and neutral washing fiber in the straws is further reduced, and more beneficial substance soluble sugar is generated. Finally, the in vitro dry matter digestibility of the corn straw yellow storage feed and the in vitro digestibility of nutrient components such as crude protein, neutral detergent fiber and acidic detergent fiber are obviously improved.
Claims (8)
1. The preparation method of the yellow-stored straw feed by the concerted fermentation of the bacterial enzyme which does not freeze at the temperature of minus 10 ℃ is characterized by comprising the following steps:
firstly, uniformly mixing corn straws and a composite type antifreezing leavening agent to obtain a raw material; the composite antifreezing leaven is prepared from a microbial agent, an active enzyme, a urea solution and sugar in a mass ratio of (1-3): 1: (1-2): (1-2) uniformly mixing; the microbial agent is prepared from bacillus subtilis subspecies, bacillus licheniformis, lactobacillus plantarum and lactobacillus johnsonii according to the mass ratio of (1.5-2.4): 1: (3.2-4.5): (4.3-5.5) in proportion; the active enzyme is prepared from cellulase, laccase, xylanase, pectinase and beta-glucanase in a mass ratio of (1-3): (1-3): (1-2): 1: (1-2) mixing; the urea solution is 30-35% by mass;
and secondly, adjusting the water content of the raw material obtained in the step one to be 50-65% by mass percent, compacting, placing the raw material into an ensiling bag, evacuating, sealing, and fermenting in dark at the temperature of more than or equal to-10 ℃ for 30-45 days to obtain the yellow silage.
2. The preparation method of the-10 ℃ non-icing fungus enzyme synergistic fermentation yellow straw silage according to claim 1, characterized in that the bacillus subtilis subspecies is purified bacillus subtilis subspecies GIM1.372 after domestication and reversion verification at low temperature of-15 ℃ to 15 ℃.
3. The preparation method of the-10 ℃ non-icing fungus enzyme synergistic fermentation silage straw feed according to claim 1 or 2, characterized in that the bacillus licheniformis is bacillus licheniformis GIM1.182 which is domesticated at a low temperature of-15 ℃ and subjected to reversion verification and purification.
4. The preparation method of the-10 ℃ non-icing bacterium-enzyme synergistic fermentation silage straw feed according to claim 1 or 2, wherein the lactobacillus plantarum is lactobacillus plantarum GIM1.648 which is domesticated at a low temperature of-15 ℃ and subjected to reversion verification and purification.
5. The preparation method of the-10 ℃ non-icing bacterium enzyme synergistic fermentation yellow straw feed according to claim 1 or 2, characterized in that the lactobacillus johnsonii is lactobacillus johnsonii GIM1.730 which is domesticated at a low temperature of-15 ℃ and subjected to reversion verification and purification.
6. The method for preparing the yellow corn stover feed through the synergistic fermentation of the bacterial enzymes and the enzymes which are not frozen at the temperature of-10 ℃ according to claim 1 or 2, wherein the corn stover is cut into sections of 1cm to 3 cm.
7. The preparation method of the yellow-stored straw feed through the bacterial enzyme synergistic fermentation at the temperature of-10 ℃ without freezing according to claim 1 or 2, wherein the mass ratio of the composite antifreezing fermentation agent to the corn straw in the step one is (0.2-0.6): 1000.
8. the method for preparing the yellow-stored straw feed through the synergistic fermentation of the bacterial enzymes which do not freeze at the temperature of-10 ℃ according to the claim 1 or 2, wherein the sugar in the step one is sucrose.
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