CN115895956A - Method for fermenting high-temperature materials by using vinasse liquid culture and application thereof - Google Patents
Method for fermenting high-temperature materials by using vinasse liquid culture and application thereof Download PDFInfo
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Abstract
The invention discloses a method for fermenting high-temperature materials by using a distiller's grain liquid culture and application thereof. The obtained fermented material can be widely applied to feed formulas of pigs, cattle, fish, laying hens and the like.
Description
Technical Field
The invention relates to the technical field of high-temperature fermentation, in particular to a method for fermenting high-temperature materials by using a grain liquid culture and application thereof.
Background
It is well known that the yield of food is greatly influenced by weather factors, and particularly when the food is influenced by extreme weather (such as cold tides, drought, hurricanes, flood disasters), the yield reduction phenomenon of the food is serious. Due to the habit of people and livestock sharing grain, the grain price for the feed raw materials is gradually increased, the feed price is high, the feed is used as a feed factory and a breeding farm of downstream enterprises, the feed cost is greatly increased due to the fact that the feed raw materials are crazy to rise, the profit of the industry is slight, even the loss is serious, and medium and small feed factories and the breeding industry suffer from inscription.
Biological extraction companies taking grain pigments as objects and traditional Chinese medicine extraction companies taking medicine and food homology as raw materials have a large amount of extract residues. According to the relevant regulations, these material residues are not intended for food for human consumption. Meanwhile, the material temperature of the material slag is high (between 75 and 82 ℃), the humidity is high (the moisture is about 40 percent or higher), and the material slag begins to decay and deteriorate 24 to 36 hours after being discharged. If the waste is treated as waste, certain resources are wasted. Because the material residues are rich in starch, protein and nutrition, the development and utilization of the materials with protein raw materials or potential energy raw materials are particularly important.
Two processing methods generally exist in the market at present, one is a puffing method: the method has high energy consumption, the cost after the expansion is greatly increased, and the method is not acceptable in feed factories or farms. The other method is a drying method: the method has high energy consumption, most of the raw materials with homology of medicine and food are rich in macromolecular structure nutrient components (such as macromolecular protein, macromolecular starch chain and the like), the macromolecular structure is not degraded, the utilization rate of animals is low, and the basic function of the raw materials of the feed is lost.
Therefore, a brand new treatment method is urgently needed, which can degrade the structures of anti-nutritional factors and macromolecules in the raw materials, is convenient to operate, has low production cost and is beneficial to market popularization.
Disclosure of Invention
The present invention is to solve the above problems, and an object of the present invention is to provide a method for fermenting a high-temperature material using a spent grain culture, which can be used for producing various feeds by treating the material by a specific method, by making full use of waste residues generated in upstream factories, and to provide an application thereof.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for fermenting high-temperature materials by using a spent grain culture comprises the following steps:
s1 selecting vinasse
Selecting glutinous sorghum and fresh rice husks, wherein the dry weight ratio is 10:2, after being respectively cooked, fully and uniformly stirring, fermenting in a wine cellar for a long time after uniformly stirring, and obtaining vinasse after nine times of cooking, eight times of fermentation and seven times of wine distillation in sequence;
s2 preparation of distillers' grains liquid
Taking the vinasse obtained in the step S1 and sterile water according to a weight ratio of 1-2: 2-5 (preferably 1:3) are added into a fermentation tank to be fully and uniformly mixed, then the mixture of the corn and the soybean meal is added to be fully and uniformly mixed, 5mol/L NaOH is used for adjusting the pH value to 7.0-7.4, then, the mixture is stirred while being aerated under the condition of 45-50 ℃, the aeration rate is 1.2-2.8 Wm, the fermentation is carried out, the culture is carried out for 2-8 hours, and the fermentation is stopped after ester fragrance appears;
s3 extraction of fermented residue liquid
1, filtering for the first time: filtering the fermentation residue liquid obtained in the step S2 by using a 30-40-mesh sterile screen, and leaving a suspension obtained by filtering;
and 2, filtering: filtering the suspension by using a 60-80-mesh sterile screen to obtain secondary filtered suspension, and storing the secondary filtered suspension in a container for later use, wherein the obtained suspension contains various strains such as bacillus subtilis, bacillus licheniformis, bacillus coagulans, mucor and saccharomycetes;
s4 culture and propagation of mixed strains
Culturing: inoculating the suspension obtained in the step S3 into a culture medium for culture;
expanding culture: inoculating the cultured mixed strain into a culture solution, and culturing for 24-72 h under the aeration condition of 1.2-1.8 wm;
s5 fermentation of high-temperature materials
And (3) inoculating the mixed strain obtained by the expanding culture in the step (S4) into the high-temperature material, wherein the inoculation temperature is 63-68 ℃, and the inoculation amount of the mixed strain in each ton of material is 2-5L.
Preferably, the water content of the distiller' S grains in the step S1 is 60-70 wt%, the crude protein content (calculated by dry matter) is 12-15 wt%, the crude fiber content (calculated by dry matter) is 18-22 wt%, and the pH value is 4.2-4.7.
Preferably, in the step S2, the corn flour and the non-shell soybean meal with the mesh number of 60-80 are mixed according to the dry weight ratio of 4-2: 1-2 (preferably 3:1), and after fully mixing, the adding amount is 5-8 wt% of the material in the fermentation tank.
Preferably, the culturing step in step S4 is specifically: (1) Sucking 0.5-1.5 mL of suspension by using a sterile suction pipe, adding the suspension into a sterilized culture plate, pouring 10-15 mL of a sterilized liquid culture medium at 50-55 ℃ into the culture plate, uniformly mixing, pouring the mixture into a constant-temperature incubator at 45 ℃ after solidification, and culturing for 48-72 h to obtain the mixed strain.
Preferably, the culture medium is prepared by the following method: accurately weighing 2-4 g of tryptone, 1-2 g of yeast extract, 2-4 g of NaCl and 3-6 g of agar powder, adding 200-400 ml of sterile water for dissolving, adding 20-40 ml of 5mol/L NaOH for uniformly mixing, adjusting the pH value to 7.0-7.4, and carrying out autoclaving at 121 ℃ for 20min.
Preferably, the culture solution in step S4 is prepared by the following method: accurately weighing 500-1000 g of tryptone, 250-500 g of yeast extract and 500-1000 g of NaCl, adding 50-100L of sterile water, dissolving, adding 5-10L of 5mol/L NaOH, uniformly mixing, adjusting the pH value to 7.0-7.4, and sterilizing at 121 ℃ under high pressure for 20min.
Preferably, the mixed strain obtained by the expanding culture in the step S4 has the bacterial content of 40 to 60 multiplied by 10 8 cfu/g。
Preferably, the step S5 further comprises the steps of filling the inoculated high-temperature material into a bag with a breathing membrane for heat sealing, starting fermentation at the ambient temperature of 52-60 ℃ (the fermentation time is 3-4 days in summer, 7-10 days in winter and 5-6 days in spring and autumn), and after the fermentation is finished, the number of viable bacteria in the fermented material is 0.15-0.3 multiplied by 10 8 cfu/g, pH value is 4.7-5.2.
Preferably, the high-temperature material is tartary buckwheat slag, the content of crude protein of the high-temperature material is 12.1% -15.5%, and the content of water is 35% -42%.
Another object of the present invention is to provide a feed prepared from the fermented product prepared by the above method, wherein the feed is added to each animal feed in the following amounts:
the invention has the beneficial effects that:
the method is used for treating the nontoxic and harmless biological leftovers and agricultural and sideline product leftovers with high temperature, high humidity, rich protein and high starch rate, and is also suitable for treating the nontoxic and harmless biological leftovers and agricultural and sideline product leftovers with normal temperature, high humidity, rich protein and high starch rate. The number of viable bacteria in the fermented material is 0.15-0.3 multiplied by 10 8 cfu/g, pH value of 4.7-5.2, and is suitable for preparing feed for pig, cattle, fish, laying hen, etc. Therefore, the invention has the following advantages:
1. the feed resource is developed, and a new path is found for relieving the feed resource deficiency;
2. the probiotics can generate various enzymes and degrade macromolecules in the raw materials, so that the digestibility of the feed can be improved, and the biological performance of animals is improved;
3. the probiotics can degrade macromolecular protein into polypeptide, so that the immunocompetence of an animal body is improved, the generated antibacterial peptide can kill harmful bacteria, and a different method is provided for antibiotic-free breeding;
4. beneficial bacteria are discharged out of the body along with excrement, so that peculiar smell harmful substances such as ammonia nitrogen, indole and the like can be continuously degraded in vitro, and the effect of purifying the culture environment is achieved.
Drawings
FIG. 1 is a schematic diagram of a colony of a mixed strain cultured according to the present invention;
FIG. 2 is a schematic view of fermentation of a high temperature material according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a colony cultured from a fermented material prepared by an embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all 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 1
The method for fermenting high-temperature materials by using the spent grain culture provided by the embodiment comprises the following steps:
s1 selection of distillers grains
Selecting glutinous sorghum and fresh rice husks, wherein the dry weight ratio of the glutinous sorghum to the fresh rice husks is 10:2, after each boiling, fully stirring evenly, fermenting in a cellar for a long time after even stirring, and sequentially carrying out nine times of boiling, eight times of fermenting and seven times of distilling, wherein the finally obtained vinasse has the water content of 68 percent, the crude protein content (calculated by dry matters) of 14.1 percent, the crude fiber content (calculated by dry matters) of 18.2 percent and the pH value of 4.4.
S2 preparation of distillers' grains liquid
Taking the fresh vinasse and sterile water, and mixing the fresh vinasse and the sterile water according to the proportion of 1: and (3) mixing the materials in a fermentation tank, and mixing the materials uniformly, wherein the weight ratio of corn flour and shelled soybean meal is 3:1, fully and uniformly mixing, and mixing the mixture of the distilled grain water and the mixture of the corn bean pulp powder according to the weight ratio of 100:7, adding the mixture into a fermentation tank, fully and uniformly mixing, adjusting the pH value to 7.2 by using 5mol/L NaOH, then, fermenting at 46 ℃ under the condition of aeration and stirring at the aeration rate of 2wm, and culturing for 8 hours until a strong ester fragrance appears.
S3 extraction of fermented liquid
Filtration for the 1 st time: filtering the above residue solution with 40 mesh sterile screen, collecting the suspension, and removing the solid on the screen
And 2, filtering: filtering the suspension with 80 mesh sterile screen to obtain filtered suspension, and storing in a container to obtain suspension containing Bacillus subtilis, bacillus licheniformis, bacillus coagulans, mucor, yeast, etc.
Carrying out verification culture on the obtained fermentation residue liquid:
sucking 1.5mL of the above fermented liquid with a sterile pipette, adding into a sterilized culture plate, adding 15mL of a sterilized culture medium at 55 deg.C into the culture plate, and mixing. After solidification, the mixture is placed in a constant temperature incubator at 45 ℃ upside down for 64h, and mixed bacterial colonies in the lees culture solution grow on the culture dish, as shown in figure 1.
Preparing 300mL of liquid culture solution, accurately weighing 3g of tryptone, 1g of yeast extract and 3g of NaCl, adding 270mL of sterile water for dissolving, adding 30mL of 5mol/L NaOH, uniformly mixing, adjusting the pH value to 7.2, and autoclaving at 121 ℃ for 20min.
Dividing the liquid culture solution into five parts, each 50mL, inoculating composite bacteria to 2 × 10 8 cfu/mL, five inoculated bacterial suspension portions were numbered A, B, C, D, E, respectively. With a as the standard. Placing in a refrigerator at 2 deg.C to prevent the growth of bacteria therein; B. c, D, E minThe culture is carried out under the constant temperature conditions of 55 ℃, 60 ℃, 65 ℃ and 70 ℃ for 72 hours, B, C, D, E culture solution is respectively taken to pass through a color comparator and a standard solution A for turbidity comparison, and then B, C, D, E is mutually turbidity compared through the color comparator, B, C, D, E turbidity is deeper than A, and E turbidity is lower than B, C, D, which shows that B, C, D, E can grow under the respective corresponding conditions, but E grows under the condition of 70 ℃, but the growth speed is worse than B, C, D.
S4 expanding culture of mixed strains
Preparation of 100L liquid Medium: 100g of tryptone, 500g of yeast extract and 1000g of NaCl, 90L of sterile water is added, 10L of 5mol/L NaOH is added after dissolution, and the pH value is adjusted to 7.2. Autoclaving at 121 deg.C for 20min.
The mixed culture of the above strains was inoculated into 100L of culture medium, and cultured at an aeration rate of 1.6wm for 72 hours.
S5 fermentation of high-temperature materials
In the embodiment, the medicinal and edible tartary buckwheat residues are taken as examples: the temperature of the tartary buckwheat residue is 78 ℃, the content of crude protein is 12.1 percent, and the water content is 35 percent. The bacterial content of each ton of tartary buckwheat slag is 60 multiplied by 10 8 5L cfu/g composite strain, the inoculation temperature is 65 ℃, and the mixture is fully and evenly stirred in an inoculator. Then put into bags with breathing films for heat sealing, and then stacked into five layers, at which the temperature between each layer of bags is 55 ℃, as shown in figure 2. When the temperature is lowered to the environmental temperature, the fermentation is mature, and the fermented material is sampled and tested for viable count and pH, respectively viable count of 0.3 multiplied by 10 8 cfu/g, and the cultured colonies are shown in FIG. 3, and the pH is 4.8.
Example 2
In this example, 60 to 90kg of lean-type growing-finishing pig feed was prepared using the fermented material prepared in example 1. The formulation of the feed is shown in table 1.
TABLE 1
| Formulation(s) | Control group | Test group |
| Corn (corn) | 54 | 54 |
| Wheat (Triticum aestivum L.) | 30 (14% moisture) | — |
| Example 1 fermentation Material | — | 30 (28% moisture) |
| Bean pulp | 12 | 12 |
| Premix compound | 4 (adding wheat enzyme) | 4 (without wheat enzyme) |
Note: the premix comprises trace mineral, major mineral, compound vitamins, amino acids, compound special wheat enzyme (xylanase and mannase), and carrier (rice chaff of about 60 meshes). If no wheat enzyme (xylanase and mannanase) is added into the premix, the wheat is difficult to digest and absorb by animals.
40 growing-finishing pigs are selected and randomly divided into two groups, wherein each group comprises 20 pigs, one group adopts the feed of the control group in the table 1, the other group adopts the feed of the test group, and the feeding experiment is carried out for 21 days, and the results are as follows:
TABLE 2
| Control group | Test group | |
| Pig head number (only) | 20 | 20 |
| The average initial weight (Kg), | 63.2 | 63..8 |
| days for rearing (heaven) | 21 | 21 |
| Total consumption material (Kg) | 1089 | 1063 |
| Average material consumption per head (Kg) | 54.45 | 53.15 |
| Average weight per head (Kg) | 79.7 | 81.4 |
| Average daily gain (Kg) | 0.79 | 0.84 |
| Average feed conversion ratio | 3.28 | 3.02 |
| Cough with dyspnea (head) | 2 | 0 |
| Thin paste (head) | 4 | 1 |
From the above experimental results, it can be seen that each index of the test group using the fermented material of example 1 is superior to that of the control group, and from the feed conversion ratio, one kg of meat per growth is saved by 0.26 kg of feed compared with the control group, and the daily gain of each pig is 0.05 kg more than that of the control group on average; in 21-day experiments, respiratory diseases of a control group are 10% higher, intestinal diseases are 15% higher, and the phenomenon is mainly related to the fact that fermented tartary buckwheat contains a large amount of fermented end products and beneficial bacteria. In addition, from the formulation, the experimental group adds the tartary buckwheat fermentation material under the condition of not adding the wheat enzyme, thereby greatly improving the digestibility of animals, and compared with the wheat which is not treated, the feeding effect is obvious.
Example 3
The fermented material in the embodiment 1 is applied to beef cattle feed, the eating desire of beef cattle to hay can be obviously improved, the fur is bright and smooth, 1-2 kg of the fermented material in the embodiment 1 is added into the conventional concentrate of each beef cattle every day, and the effect of obviously promoting the beef cattle to grow fat is achieved.
Example 4
When the fermented material of example 1 is applied to fish feed, particularly in the weather of 7,8,9 at high temperature, the fermented material of example 1 is added according to the proportion that the weight ratio of the complete feed is 15% when the fish feed is fed, and the feed intake of fish flocks is obviously improved after 2-3 days (the fish flocks are reluctant to leave for a long time after the fish flocks eat the feed fed by the feeder). The energy is saved in high temperature season, the water quality is well improved (the PH value is 7.5, the dissolved oxygen is 8mg/l, the ammonia nitrogen is =0.12mg/l, the transparency is 34 cm), and intestinal diseases, septicemia, fin rot and the like of fish are obviously reduced.
Example 5
When the fermented material in the example 1 is applied to the feed of the laying hens, 35% of corn and all bran in the formula are replaced by the fermented material in the example 1, and compared with the fermented material without replacement, the laying rate is increased quickly, the high yield peak value is maintained for 12 days more, the death and culling rate is reduced by about 3%, and the breakage rate of eggs is also reduced by 5%.
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 (10)
1. A method for fermenting high-temperature materials by using a spent grain culture is characterized by comprising the following steps:
s1 selecting vinasse
Selecting glutinous sorghum and fresh rice husks, wherein the dry weight ratio is 10:2, after each steaming and boiling, fully and evenly stirring, and fermenting in a cellar for a long time after evenly stirring. Carrying out nine times of stewing, eight times of fermentation and seven times of wine distillation in sequence to obtain vinasse;
s2 preparation of distillers' grains liquid
Taking the vinasse obtained in the step S1 and sterile water according to a weight ratio of 1-2: 2-5 (preferably 1:3) are added into a fermentation tank to be fully and uniformly mixed, then the mixture of the corn and the soybean meal is added to be fully and uniformly mixed, 5mol/L NaOH is used for adjusting the pH value to 7.0-7.4, then, the mixture is stirred while being aerated under the condition of 45-50 ℃, the aeration rate is 1.2-2.8 Wm, the fermentation is carried out, the culture is carried out for 2-8 hours, and the fermentation is stopped after ester fragrance appears;
s3 extraction of fermented residue liquid
Filtration for the 1 st time: filtering the fermentation residue liquid obtained in the step S2 by using a 30-40-mesh sterile screen, and leaving a suspension obtained by filtering;
and (3) filtering for the 2 nd time: filtering the suspension by using a 60-80-mesh sterile screen to obtain secondary filtered suspension, and storing the secondary filtered suspension in a container for later use, wherein the obtained suspension contains various strains such as bacillus subtilis, bacillus licheniformis, bacillus coagulans, mucor and saccharomycetes;
s4 culture and propagation of mixed strains
Culturing: inoculating the suspension obtained in the step S3 into a culture medium for culture;
expanding culture: inoculating the cultured mixed strain into a culture solution, and culturing for 24-72 h under the aeration condition of 1.2-1.8 wm;
s5 fermentation of high-temperature materials
And (3) inoculating the mixed strain obtained by the expanding culture in the step (S4) into the high-temperature material, wherein the inoculation temperature is 63-68 ℃, and the inoculation amount of the mixed strain in each ton of material is 2-5L.
2. The method according to claim 1, wherein the water content of the distiller' S grains in the step S1 is 60-70 wt%, the crude protein content (on a dry matter basis) is 12-15 wt%, the crude fiber content (on a dry matter basis) is 18-22 wt%, and the pH value is 4.2-4.7.
3. The method as claimed in claim 1, wherein the corn flour and the non-shell soybean meal with the mesh number of 60-80 in the step S2 are mixed fully and uniformly according to the dry weight ratio of 4-2:1-2 (preferably 3:1) and the adding amount is 5-8 wt% of the material in the fermentation tank.
4. The method according to claim 1, wherein the culturing step in step S4 is specifically: (1) Sucking 0.5-1.5 mL of suspension by using a sterile suction pipe, adding the suspension into a sterilized culture plate, pouring 10-15 mL of a sterilized culture medium at 50-55 ℃ into the culture plate, uniformly mixing, pouring the mixture into a constant-temperature incubator at 45 ℃ after solidification, and culturing for 48-72 h to obtain the mixed strain.
5. The method of claim 4, wherein the culture medium is prepared by: accurately weighing 2-4 g of tryptone, 1-2 g of yeast extract and 2-4 g of NaCl, adding 3-6 g of agar powder, adding 200-400 ml of sterile water for dissolving, adding 20-40 ml of 5mol/L NaOH for uniformly mixing, adjusting the pH value to 7.0-7.4, and sterilizing at 121 ℃ for 20min under high pressure.
6. The method according to claim 1, wherein the culture solution in step S4 is prepared by the following method: accurately weighing 500-1000 g of tryptone, 250-500 g of yeast extract and 500-1000 g of NaCl, adding 50-100L of sterile water, dissolving, adding 5-10L of 5mol/L NaOH, uniformly mixing, adjusting the pH value to 7.0-7.4, and sterilizing at 121 ℃ under high pressure for 20min.
7. The method according to claim 1, wherein the mixed bacterial strain obtained by the expanding culture in step S4 has a bacterial content of 40-60 x 10 8 cfu/g。
8. The method of claim 1, wherein step S5 further comprises the steps of placing the inoculated high-temperature material into a bag with a breathing membrane for heat sealing, starting fermentation at an ambient temperature of 52-60 ℃ (the fermentation time is 3-4 days in summer, 7-10 days in winter, and 5-6 days in spring and autumn), and after the fermentation is finished, the number of viable bacteria in the fermented material is 0.15-0.3 x 10 8 cfu/g, pH value is 4.7-5.2.
9. The method according to claim 1, wherein the high-temperature material is tartary buckwheat slag, the content of crude protein of the tartary buckwheat slag is 12.1% -15.5%, and the content of water is 35% -42%.
10. An animal feed comprising the fermented material prepared by the method of any one of claims 1 to 9 in the following amounts:
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